CN101258153A - Cysteine protease inhibitors - Google Patents

Abstract

The present invention provides a compound of the formula (II) wherein one of R<1> and R<2> is halo and the other is H or halo; R<3> is -C1-C5 straight or branched chain, optionally fluorinated, alkyl or -CH2CR<5>C3-C4-Cycloalkyl; R<4> is H; R<5> is H, C1-C2 alkyl, C1-C2 haloalkyl, hydroxyl, OC1-C2alkyl, fluoro; R<6> is a stable, optionally substituted, monocyclic or bicyclic, carbocycle or hetorocycle wherein the or each ring has 4, 5 or 6 ring atoms and 0 to 3 hetero atoms selected from S, O and N; Rb is haloalkyl; Rc is H or C1-C4 alkyl; and pharmaceutically acceptable salts, hydrates or N-oxides thereof have utility in the treatment of disorders characterised by inappropriate expression or activation of cathepsin K, such as osteoporosis, osteoarthritis, rheumatoid arthritis or bone metastases.

Description

Cystatin

Technical field

The present invention relates to cystatin, particularly the inhibitor of those papoid superfamilies.The invention provides and be used for coming from the prevention of the unbalance disease of physiology proteolytic enzyme (such as cathepsin K) or the compounds for the treatment of.

The description of association area

The papoid superfamily of L-Cysteine HCL Anhydrous is distributed widely in the multiple species that comprise Mammals, invertebrates, protozoon, plant and bacterium.With multiple mammalian tissues proteolytic enzyme, comprise that cathepsin B, F, H, K, L, O and S belong to this superfamily, and their active inappropriate adjustment and multiple Metabolic disorder have complicated getting in touch, and comprise sacroiliitis, muscular dystrophy, inflammation, glomerulonephritis and tumour intrusion.Comprise gum porphin beautiful jade Zymomonas mobilis (bacterial gingipains), malarial falcipains I, II, III and subordinate thereof and derive from the L-Cysteine HCL Anhydrous of Pneumocystis carinii, schizotrypanum cruzi and trypanosoma bocagei, Crithidiafusiculata, schistosomicide species such as the pathogenic kethepsin of enzyme.

The inappropriate adjustment of cathepsin K has complicated getting in touch with multiple disease, comprises the hypercalcemia and the metabolic skeletal diseases of osteoporosis, gum disease (such as gingivitis and periodontitis), Paget's disease, malignant tumour.In view of the elevated levels in the chondroclast of osteoarthritis synovial membrane, cathepsin K and various features are that the disease of excessive cartilage or matrix degraded has complicated getting in touch, such as osteoarthritis and rheumatoid arthritis.Metastatic tumour cell is general expresses the proteolytic ferment of matrix around the high-caliber degraded and thus, can help to treat tumour to the restraining effect of cathepsin K and form.

International Patent Application WO 02057270 discloses formula I compound:

Wherein UVWXY is broadly corresponding to the P3 and the P2 of dipeptides cystatin, Z especially be O, S, methylene radical or-NR-, R ¹Be alkyl, alkylaryl or the like and P1 and the Q1 methylene radical of respectively doing for oneself, optionally replaced by multiple carbochain and cyclic group.Described compound is declared can be used for the treatment of such as leishmanial protozoal infections.Wherein there is not specifically to disclose the haloalkyl isostere of P2/P3 amido linkage.

We find now, introduce halogen atom at specific ring position and connect the effective inhibitor that produces cathepsin K in conjunction with halogenation P3/P2.

The invention summary

According to the present invention, provide formula II compound:

Wherein

R ¹And R ²In one of be that halogen and another are H or halogen;

R ³Be optional fluorizated-C ₁-C ₅Straight chain or branched-chain alkyl, perhaps CH ₂CR ⁵C ₃-C ₄-cycloalkyl;

R ⁴Be H;

R ⁵Be H, C ₁-C ₂Alkyl, C ₁-C ₂Haloalkyl, hydroxyl, OC ₁-C ₂Alkyl, fluorine;

R ⁶Be monocycle stable, optional replacement or two ring carbocyclic ring or heterocycles, described or each ring has 4,5 or 6 annular atomses and 0～3 substituting group that is selected from the heteroatoms of S, O and N and wherein chooses wantonly comprises that 1～3 is selected from R ₇The member;

R ₇Be independently selected from halogen, oxo, nitrile, nitro, C ₁-C ₄Alkyl ,-XNRdRe ,-XNReR ⁸,-NReXR ⁸, NH ₂CO-, X-R ⁸, X-O-R ⁸, O-X-R ⁸, X-C (=O) R ⁸, X-(C=O) NRdR ⁸, X-NReC (=O) R ⁸, X-NHSO _mR ⁸, X-S (=O) _mR ⁸, X-C (=O) OR ⁸, X-NReC (=O) OR ⁸

R ⁸Be H, C independently ₁-C ₄Alkyl, C ₃-C ₆Cycloalkyl, pyrrolidyl, piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, indolinyl, pyranyl, sulfo-pyranyl, furyl, thienyl, pyrryl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, indyl, phenyl, they are chosen wantonly separately and are selected from R by maximum 3 ⁹The member replace;

R ⁹Be independently selected from hydroxyl, XR ¹⁰,-XNRdRe ,-XNReR ¹⁰,-NReC ₁-C ₄Alkyl R ¹⁰, cyano group ,-S (=O) _mRe, carboxyl, oxo, C ₁-C ₄Alkyl, C ₁-C ₄Haloalkyl, C ₁-C ₄-alkoxyl group, C ₁-C ₄Alkyloyl, formamyl;

R ¹⁰Be C ₃-C ₆Cycloalkyl, pyrrolidyl, piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, indolinyl, pyranyl, sulfo-pyranyl, furyl, thienyl, pyrryl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, indyl, phenyl, they are arbitrary by C ₁-C ₄Alkyl, halogen, hydroxyl, C ₁-C ₄Alkoxyl group replaces;

X is associative key or C independently ₁-C ₄Alkylidene group;

Ra is H, C independently ₁-C ₄Alkyl or CH ₃C (=O);

Rb is C ₁-C ₄Haloalkyl;

Rc is H, C ₁-C ₄Alkyl;

Rd is H, C independently ₁-C ₄Alkyl or CH ₃C (=O);

Re is H, C independently ₁-C ₄Alkyl; Perhaps

Rd forms optional by R with the N atom that Re is connected with them altogether ⁹The morpholine, piperidines, piperazine or the pyrrolidine ring that replace;

M is 0,1 or 2 independently;

Perhaps its pharmacy acceptable salt, hydrate or N-oxide compound.

Do not wish by theory it to be fettered by any way or tentative connection mode is belonged to concrete variable, P1, P2 and P3 in this application provide for the purpose of facility, they have their conventional sense, the expression inhibitor is considered to those parts of the described enzyme S1 of difference filling, S2 and S3 sublocus, and wherein contiguous broken site of S1 and S3 are away from broken site.

The stereochemistry of preferred P1 group is as indicated in the following part-structure:

Preferred R ¹And/or R ²Halogen be chlorine, and most preferably be fluorine.Current preferred R ²Be halogen, particularly fluorine, and R ¹Be H, but the present invention extends to wherein R ¹Be halogen, particularly F, and R ²Be H or R ¹And R ²Respectively the do for oneself compound of F.

Should be appreciated that the P1 group can exist with alterative version, such as

And the present invention extends to all described alterative version.

The stereochemistry of preferred P2 group is corresponding to the L-amino acid as indicated in following part-structure:

But the present invention also extends to the D-isomer.

The present invention also is included in all isomer and the enantiomer on other chiral centre.

Current preferred P2 group comprises wherein R ⁴Be H and R wherein ³For isobutyl-or all the tertiary butyl (promptly-CH ₂C (CH ₃) ₃) those P2 groups, as follows:

Work as R ⁴During for H, R ³Other embodiment comprise those with following part-structure:

R wherein ⁵As defined above.

Desirably, R ⁵Be H, define the cyclobutylmethyl base side chain on the P2 thus.

R ⁵Typical value comprise methyl, hydroxyl, fluoro methyl, difluoromethyl or trifluoromethyl: in view of the above, the preferred value of P2 side chain comprises,

Particularly reflect L amino acid whose those.

Current preferred P2 group comprises

Current, show that preferably the Ra in formula II is H.

The haloalkyl of preferred Rb comprises halogenated methyl, such as fluoro methyl, difluoromethyl and preferred trifluoromethyl.

Usually Rc is H and R ⁶Be the independent ring system shown in following part-structure:

Purpose for the purpose of illustration wherein, R ⁶Illustration be that substituted-phenyl and Rb are trifluoromethyl:

Preferred The compounds of this invention is included in the S steric configuration that has high optical isomer purity on the carbon that has haloalkyl Rb, such as greater than 80%, is preferably greater than 95%, such as greater than 97%.Following part-structure represents that Rb is that trifluoromethyl and Rc are the typical S-enantiomer of H:

Be back to formula II now, general R ⁶For having 5 or the monocycle of 6 annular atomses particularly, perhaps comprise two ring structures that are fused to 6 yuan of rings on 4,5 or 6 yuan of rings.

General R ⁶Group comprises saturated or undersaturated heterocycle or saturated or undersaturated carbocyclic ring, and they are chosen wantonly separately and are substituted as mentioned above.Illustrative variable comprises C _3-8Cycloalkyl, phenyl, benzyl, tetralyl, indenyl, indanyl and heterocyclic radical, such as the azepan base, azocanyl, pyrrolidyl, piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, indolinyl, pyranyl, THP trtrahydropyranyl, tetrahydro thiapyran base, the sulfo-pyranyl, furyl, tetrahydrofuran base, thienyl, pyrryl oxazolyl isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, tetrazyl, pyrazolyl, indyl, benzofuryl, benzothienyl, benzimidazolyl-, benzothiazolyl benzoxazolyl, the benzoisoxazole base, quinolyl, tetrahydric quinoline group, isoquinolyl, tetrahydro isoquinolyl, quinazolyl, tetrahydro quinazoline base and quinoxalinyl, its arbitrary can being substituted as mentioned above.

Thus, described saturated heterocyclic comprises such as following group, pyrrolinyl, pyrrolidyl, pyrazolinyl, pyrazolidyl, piperidyl, morpholinyl, thio-morpholinyl, pyranyl, the sulfo-pyranyl, piperazinyl, indolinyl, azetidinyl, THP trtrahydropyranyl, tetrahydro thiapyran base, tetrahydrofuran base, the hexahydropyrimidine base, the hexahydro-pyridazine base, 1,4,5,6-tetrahydro-pyrimidine base amine dihydro-oxazole base, 1,2-thiazinyl-1, the 1-dioxide, 1,2,6-thiadiazine base-1, the 1-dioxide, isothiazoline base-1,1-dioxide and imidazolinyl-2, the 4-diketone, unsaturated heterocycle comprises following group simultaneously, such as furyl, thienyl, pyrryl oxazolyl, thiazolyl, imidazolyl, pyrazolyl isoxazolyl, isothiazolyl oxadiazole base, triazolyl, tetrazyl, thiadiazolyl group, pyridyl, pyridazinyl, pyrimidyl, pyrazinyl, the indolizine base, indyl, pseudoindolyl.In various situations, described heterocycle can with the benzyl ring condensation, thereby form bicyclic system.

Preferred monocycle R ⁶Group comprises substituted pyridinyl, substituted pyrimidyl, substituted-phenyl, the phenyl that is replaced by cyclic group particularly, described cyclic group is such as tetramethyleneimine-1-base, piperidines-1-base, 4-methyl piperidine-1-base, 4-(piperidines-3-ylmethyl)-piperidines-1-base, morpholine-4-base, 4-methylpiperazine-1-base, 2-morpholine-4-base-ethylamino, 2-morpholine-4-base-oxyethyl group, 1-pyridine-2-ylmethyl amino, piperazine-1-base, piperidin-4-yl or N-piperazinyl, replaced by Ra N-, perhaps piperidines-1-the base that is replaced by-NRaRb4-.Phenyl R ⁶Desirably 3 or 4 (to or a position) be substituted, for example replaced by described cyclic group.

Monocycle R ⁶The another kind of cyclic substituents of (such as phenyl) comprises aryl (such as phenyl) or 5 or 6 yuan of heteroaryls (such as thiophene, furyl, triazole, thiazole, diazole, pyrazoles or tetramethyleneimine).About this point, preferred cyclic substituents comprises thiazol-2-yl, pyridin-3-yl and particularly pyridine-2-base, thiophene-2-base or thiazole-5-base.This cyclic substituents (is R ⁷) generally directly be connected to described R ⁶On the group (being that X is an associative key), but for example can also comprise the amine spacer (such as-NH-,-N (Me) ,-CH ₂NH ,-CH ₂N (Me)-), C ₁-C ₃The alkyl spacer (such as-CH ₂-) or C ₁-C ₃-alkoxyl group spacer (such as oxyethyl group).

In the last period, R ⁶Any cyclic substituents can be as mentioned above by R ¹⁰Replace.For example, such as the heterocycle R of thiazolyl ⁷Group can be by C ₁-C ₄Alkyl (such as methyl) replaces.

Preferably in preceding two sections, R ⁶Any cyclic substituents cyclic group (such as piperidines, piperazine or morpholine) that can self be generally the saturated heterocyclic group replace (that is R, ⁷Comprise R ⁹Part), described saturated cyclic group is optional to be substituted, for example by C ₁-C ₃Alkyl, fluorine, difluoro, C ₁-C ₃Alkoxyl group or C ₁-C ₃Alkoxy C ₁-C ₃Alkyl replaces.As R ⁷Provide in the definition, this saturated cyclic group (is R ⁹) can pass through X (C for example ₁-C ₃Alkyl), amine (for example-NH-), acid amides, amine sulphonyl or the like and R ⁶Group is isolated, but general direct keyed jointing or through the methylene radical keyed jointing.

Representative R corresponding to the last period ⁹Group comprises heterocycle, such as tetramethyleneimine-1-base, piperidines-1-base, 4-methyl piperidine-1-base, 4-(piperidines-3-ylmethyl)-piperidines-1-base, morpholine-4-base, 4-methylpiperazine-1-base, 2-morpholine-4-base-ethylamino, 2-morpholine-4-base-oxyethyl group, 1-pyridine-2-ylmethyl amino, piperazine-1-base, piperidin-4-yl or N-piperazinyl, replaced by Ra N-, perhaps piperidines-1-the base that is replaced by-NRaRb 4-.

Current preferred R ⁹Substituting group comprises piperazine-4-base (such as 4-methyl-piperazine-4-base or 4-methoxy ethyl-piperazine-4-yl) that 4-replaces, optional piperidines-1-ylmethyl or the morpholinyl methyl that is replaced by fluorine or difluoro.

Monocycle R ⁶Other preferred substituents of (such as phenyl) comprises-NRaRb ,-CH ₂NRaRb, C ₁-C ₄Straight chain or branched-chain alkyl or-O-R ⁹

Monocycle R ⁶Other preferred substituents of (such as phenyl) comprises the group that contains non-basic moiety, such as halogen, hydroxyl, carboxyl, C ₁-C ₄Haloalkyl, C ₁-C ₄Alkoxyl group and following formula group:

-S (=O) _mC ₁-C ₄Alkyl ,-S (=O) _mC ₃-C ₆Cycloalkyl or have the cycloalkyl that the formamyl of following part-structure replaces:

Wherein Rg is H, C ₁-C ₄Alkyl or cyclopropyl, Rh is H, C ₁-C ₄Alkyl; Perhaps Rg limits tetramethyleneimine, morpholine, piperidines, piperazine or N methyl piperazine altogether with the N atom that Rh is connected with them.

Representational R ⁶Group comprises:

Other representational R ⁶Group comprises:

Particularly

Wherein Rq and Rq ' are independently selected from H, C ₁-C ₄Alkyl or C ₁-C ₄Alkyloyl or limit unsaturated 5-7 unit ring together, such as piperidines, piperazine or morpholine, can so by corresponding to R ¹⁰Group replace C particularly ₁-C ₄Alkyl, fluorine or difluoro.

Current preferred R ⁶Group comprises

For R ⁶, representational bicyclic groups comprises naphthyl, particularly naphthalene-2-base;

Benzo [1,3] dioxa cyclopentenyl, particularly benzo [1,3] dioxole-5-base, benzofuryl, particularly cumarone-2-base and particularly C ₁-C ₆The benzofuryl that alkoxyl group replaces, more especially 5-(2-piperazine-4-carboxylic acid tert-butyl ester oxyethyl group) cumarone-2-base, 5-(2-morpholine-4-base-oxyethyl group)-cumarone-2-base, 5-(2-piperazine-1-base-oxyethyl group) cumarone-2-base, 5-(2-cyclohexyl-oxyethyl group)-cumarone-2-base; 7-methoxyl group-cumarone-2-base, 5-methoxyl group-cumarone-2-base, 5, the benzofuryl that 6-dimethoxy-cumarone-2-base, particularly halogen replace, more especially 5-fluoro-cumarone-2-base, 5,6-two fluoro-cumarone-2-base, particularly C ₁-C ₆Benzofuryl, the most particularly 3-methyl-cumarone-2-base that alkyl replaces; Benzo [b] thienyl, particularly benzo [b] thiophene-2-base; C particularly ₁-C ₆Benzo [b] thienyl that alkoxyl group replaces, more especially 5,6-dimethoxy-benzo [b] thiophene-2-base, quinolyl, particularly quinoline-2-base, quinoline-3-base, quinolyl-4, quinoline-6-base and quinoline-S-base; Quinoxalinyl, particularly quinoxaline-2-base; 1,8-naphthyridinyl, particularly 1,8-naphthyridines-2-base; Indyl, particularly indoles-2-base, particularly indoles-6-base, indoles-5-base, particularly C ₁-C ₆The indyl that alkyl replaces, more especially N-skatole-2-base; Furo [3,2-b] pyridyl, particularly furo [3,2-b] pyridine-2-base, and C ₁-C ₆Furo [3,2-b] pyridyl, particularly 3-methyl-furo [3,2-b] pyridine-2-base that-alkyl replaces; Thieno-[3,2-b] thiophene, particularly thieno-[3,2-b] thiophene-2-base, more especially C ₁-C ₆Thieno-[3,2-b] thiophene-2-base that alkyl replaces, more especially the 5-tertiary butyl-3 methyl thiophene [3,2-b] thiophene-2-base also.

Favourable R ⁶Group comprises two rings, and such as naphthyl, quinoline acyl group, benzofuryl, benzothienyl, indyl and indolinyl, special wherein connector is connected on 2 of ring.Two ring R ⁶Favourable substituting group comprise tetramethyleneimine-1-base, piperidines-1-base, 4-methyl piperidine-1-base, 4-(piperidines-3-ylmethyl)-piperidines-1-base, morpholine-4-base, 4-methylpiperazine-1-base, 2-morpholine-4-base-ethylamino, 2-morpholine-4-base-oxyethyl group, 1-pyridine-2-ylmethyl amino, piperazine-1-base, piperidin-4-yl or N-piperazinyl, replaced by Ra N-, perhaps piperidines-1-the base that is replaced by-NRaRb 4-.Particularly preferred substituting group, particularly the substituting group in conjunction with benzofuryl comprises 2-morpholine-4-base-oxyethyl group and N-methyl-piperidin-4-yl oxygen base and the substituting group as giving a definition.

Current preferred two ring R ⁶Group is optional benzothiazole or benzofuryl or the benzoxazolyl that replaces, and comprises that wherein substituting group is-OR ⁹Perhaps-NRbR ⁹Those.For example, favourable R ⁶Group comprises cumarone-2-base, be not substituted or at 3 by C ₁-C ₄Alkyl (such as methyl) or C ₁-C ₄Haloalkyl (such as trifluoromethyl) replaces and/or is replaced by saturated heterocyclic (such as piperidines, piperazine or morpholine) at 5, and described heterocycle is optional by C ₁-C ₃Alkyl replaces and/or leads to peroxy, methoxyl group or oxyethyl group and benzofuryl interval.Thus, particularly advantageous benzofuryl R ⁶Group comprises:

Recoverable II, usually:

X is generally methylene radical or particularly associative key.

Wherein n is 4 C ₁-C _nAlkyl self or such as C ₁-C ₄In the compound expression formula of alkoxyl group, comprise methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, cyclopropyl and methyl cyclopropyl or the like, for other n value, expansion in a similar fashion.For example, C ₅Alkyl comprises the equal tertiary butyl (CH ₂C (CH ₃) ₃).

Halogen or halogen comprise bromine, chlorine and particularly fluorine.

Haloalkyl is meant that at least one carbon atom wherein has the alkyl as defined above of 1～3 halogen atom, preferred fluorine atom.Representational haloalkyl comprises fluoro methyl, difluoromethyl, trifluoromethyl, 2-fluoro ethyl, 2,2-two fluoro ethyls and 2,2,2-trifluoroethyl or the like.

Favourable The compounds of this invention comprises that those pass through the arrangement of selecting P3, P2 and P1 member to form independently from each form A, B and C:

Table A P1 group

Table BP2 group

Table CP3 group

Another aspect of the present invention comprises pharmaceutical composition, comprises compound and pharmaceutically acceptable carrier or thinner as defined above.

Another aspect of the present invention relates to compound as defined above and is used for the treatment of purposes in the medicine of disease of cathepsin K mediation in manufacturing, such as:

Osteoporosis,

Gum disease, such as gingivitis and periodontitis,

Paget's disease,

The hypercalcemia of malignant tumour

The metabolism skeletal diseases

Be characterised in that the disease of excessive cartilage or matrix degraded, such as osteoarthritis and rheumatoid arthritis,

Osteocarcinoma comprises that tumour forms,

Pain.

Think that the present invention has specific end use to osteoporosis, osteoarthritis, rheumatic arthritis and/or bone transfer.

The compounds of this invention can form salt, and this constitutes another aspect of the present invention.The suitable pharmacy acceptable salt of formula II compound comprises organic acid salt, carboxylate salt particularly, include but not limited to acetate, trifluoroacetate, lactic acid salt, gluconate, Citrate trianion, tartrate, maleate, malate, pantothenate, isethionate, adipate, alginate, aspartate, benzoate, butyrates, digluconate, chaulmoogric acid salt, glucoheptose salt, glycerophosphate, oxalate, enanthate, hexanoate, fumarate, nicotinate, pamoate, pectinate, 3-phenylpropionic acid salt, picrate, pivalate, propionic salt, tartrate, Lactobionate, pivolate, camphorate, undecylate and succinate, organic sulfonate is such as mesylate, esilate, the 2-isethionate, camsilate, the 2-naphthalenesulfonate, benzene sulfonate, closilate and tosilate; And inorganic acid salt, such as hydrochloride, hydrobromate, hydriodate, vitriol sulfuric acid hydrogen salt, Hemisulphate, thiocyanate-, persulphate, phosphoric acid salt and sulfonate.In some cases, can also be hydrate with formula II compound separation.Hydrate is generally by in moisture/ORGANIC SOLVENT MIXTURES, uses organic solvent such as dioxin, tetrahydrofuran (THF) or methyl alcohol to carry out recrystallization and is prepared.

The N-oxide compound of formula (I) compound can be prepared by the method that those skilled in the art know.For example, the N-oxide compound can be prepared by the following method, under about 0 ℃, at suitable inert organic solvents (for example, halohydrocarbon, such as methylene dichloride) in, with oxygenant (for example, trifluoroperacetic acid, cross toxilic acid, peroxybenzoic acid, peracetic acid or-chloroperoxybenzoic acid or the like) the not oxidised form of processing formula (I) compound.Additionally, the N-oxide compound of formula (I) compound can be prepared by the N-oxide compound of suitable raw material.

The formula of oxidised form (I) compound can be prepared in the following manner by the N-oxide compound of formula (I) compound, under 0～80 ℃, at suitable inert organic solvents (for example, acetonitrile, ethanol or moisture dioxane or the like) in, handle with reductive agent (for example, sulphur, sulfurous gas, triphenyl phosphine, lithium borohydride, sodium borohydride, phosphorus dichloride or phosphorus tribromide or the like).

Formula (II) compound can be prepared to their single steric isomer in the following manner, make the racemic mixture and the reaction of optically active resolution reagent of compound, thereby form a pair of diastereomeric compound, separate diastereomer and reclaim optically pure enantiomer.Though the fractionation of enantiomer can utilize the covalency diastereo-isomerism derivative of formula (I) compound to carry out, preferred separable title complex (for example, crystal; The salt of diastereomer).Diastereomer has different physical properties (for example, fusing point, boiling point, solvability, reactivity or the like), and can obtain delamination by utilizing these differences.Diastereomer can separate by chromatography, and for example HPLC perhaps is preferably based on deliquescent difference and separates by separation/resolution process.Then, reclaim optically pure enantiomer and resolution reagent by any hands-on approach of racemization that can not cause.Be applicable to that being described in more detail of technology that splits the steric isomer of compound from their racemic mixture can be found in Jean Jacques AndreCollet, Samuel H.Wilen, Enantiomers, Racemates and Resolutions, JohnWiley﹠amp; Sons is among the Inc. (1981).

Should be appreciated that the present invention extends to the form of release type II compound in prodrug, solvate, title complex and other body.

Though can promoting agent is individually dosed, preferably it exists as the part of pharmaceutical preparation.Described preparation will comprise promoting agent and one or more acceptable carrier/vehicle and other optional therapeutics composition of above-mentioned definition.Described carrier must be " acceptable ", its implication be with preparation in other composition compatible and harmless to its receptor.

Described preparation comprises and is applicable to rectum, intranasal, part (comprising oral cavity and hypogloeeis), vagina or parenteral (comprising subcutaneous, intramuscular, intravenously and intradermal) administration, but preferred described preparation is an oral Preparation.Described preparation can be desirably exists with the form of unit dosage form, for example tablet and slow releasing capsule, and can be prepared by the method that any pharmacy field is known.

Described method comprises the promoting agent of above-mentioned definition and carrier-bound step.Usually, pharmaceutical composition is prepared in the following manner: combine with liquid vehicle or solid carrier in small, broken bits or the two nearly activeconstituents is all even, subsequently, if necessary, it is shaped to product.The present invention includes the method for pharmaceutical compositions, comprise making formula II compound or its pharmacy acceptable salt with pharmaceutically acceptable carrier or vehicle associating or combine.If the manufacturing of pharmaceutical preparation relates to the activeconstituents of close hybrid medicine vehicle and salt form, the preferred so usually vehicle of non-alkalescence in essence that uses, promptly acid or neutral vehicle.

In the present invention, the preparation that is used for oral administration can exist for separating unit, and such as capsule, cachet or tablet, they contain the promoting agent of predetermined amount separately; Be pulvis or granula; Perhaps be solution or the suspension agent of promoting agent in liquid, aqueous or on-aqueous liquid; Perhaps be oil-in-water liquid emulsion or water-in-oil liquid emulsion and for bullet or the like.

For liquid preparations for oral administration (for example tablet and capsule), the suitable carrier of term comprises vehicle, such as ordinary excipients, for example tackiness agent, for example syrup, gum arabic, gel, sorbyl alcohol, tragakanta, polyvinylpyrrolidone (Povidone), methylcellulose gum, ethyl cellulose, Xylo-Mucine, Vltra tears, sucrose and starch; Filler and carrier, for example W-Gum, gel, lactose, sucrose, Microcrystalline Cellulose, kaolin, N.F,USP MANNITOL, Lin Suanergai, sodium-chlor and alginic acid; And lubricant, such as Magnesium Stearate, sodium stearate and other metallic stearate, Vinlub stearic acid, silicone fluid, talcum, paraffin, oils and colloided silica.Can also use sweetener, such as Mentha arvensis L. syn.M.haplocalyxBrig, wintergreen oil, cherry flavour or the like.Can desirably add tinting material, thereby make that formulation can be by identification easily.Can also carry out dressing to tablet by method well known in the art.

Tablet can be prepared by compression or mold, and is optional with one or more auxiliary agents.Tablet agent can be prepared by the promoting agent that compresses free-flowing form (such as powder or particle) in suitable equipment, optional described promoting agent and wedding agent, lubricant, inert diluent, sanitas, tensio-active agent or dispersant.The mold tablet can be prepared by the mixture of mold in suitable equipment with the wetting powder compounds of inert liquid diluent.Can choose wantonly tablet is carried out dressing or indentation and prepares, thereby the slowly-releasing or the controlled release of promoting agent are provided.

Other preparation that is applicable to oral administration comprises lozenge, wherein promoting agent is included in the spices matrix, is generally sucrose and gum arabic or tragakanta; Pastille wherein is included in promoting agent in the inertial base, such as gel and glycerine or sucrose and gum arabic; And mouth-washes, wherein promoting agent is included in the appropriate liquid carrier.

The suitable dosage of The compounds of this invention or preparation will depend on indication and patient, and can determine easily by the conventional animal test.It generally is desirable and obtainable that the dosage of (being used to suppress the physiology proteolytic enzyme of papoid superfamily) concentration in the born of the same parents of about 0.01-100 μ M is provided, and more preferably 0.01-10 μ M is such as 0.1-25 μ M.

The compounds of this invention is prepared by multiple solution and solid state chemistry process.

Compound generally is prepared as P1, the P2 of reflection the finished product inhibitor and the structural unit of P3 part.Do not wish by theory it to be fettered by any way or tentative connection mode is belonged to concrete variable, abstract concept P1, P2 and P3 in this application provide for the purpose of facility, and they have their conventional Schlecter﹠amp basically; The Berger implication, the expression inhibitor is confirmed to be those parts of the described enzyme S1 of difference filling, S2 and S3 sublocus, and wherein contiguous broken site of S1 and S3 are away from broken site.No matter mode of connection how, the compound of through type I definition all is intended to be included in the scope of the present invention.

Broadly, the P1 structural unit will be N-protected-6-fluoro-3-oxo-six hydrogen-furo [3,2-b] pyrroles; P2 will be the amino acid of N-protected; P3 generally comprises capping group, and such as the 4-hetaroylpyrazol or the aroyl part that replace, the carbon connector that replaces through the Rb-haloalkyl is connected on the P2.

The single structure unit of due care can at first be prepared and subsequently they are coupled at together, i.e. P2+P1 → P2-P1.Additionally, the precursor of structural unit can be coupled at together and carry out modification in the step afterwards of synthetic inhibitor sequence.Then, structural unit, structural unit precursor or the bigger fragment of ready-formed of other desired structure can be coupled on the growing chain, for example R ³-E-P2 ^*+ P1 → R ³-E-P2-P1 or R ³-E ^*+ P2-P1 → R ³-E-P2-P1, wherein ^*The expression activated form.

The formation of peptide bond, be that coupling can utilize the standard coupling method to carry out, such as the trinitride method, mix carbon-carboxylic acid anhydride (isobutyl chlorocarbonate) method, carbodiimide (dicyclohexylcarbodiimide, DIC or water miscible carbodiimide) method, active ester p-nitrophenyl ester, N-hydroxy-butanedioic acid imino esters) method, Woodward reagent K-method, N,N'-carbonyldiimidazole method, phosphorus reagent or oxidation reduction process.Some aforesaid methods (particularly carbodiimide method) can be enhanced by adding I-hydroxybenzotriazole or 4-DMAP.These linked reactions can be carried out in solution (liquid phase) or solid phase.

More clearly, coupling step relates in the presence of coupling agent, makes a kind of free amine group dehydrogenation coupling of free carboxy and another kind of reactant of reactant, thereby forms the amido linkage that connects.The description of described coupling agent is typically found in the general textbook about chemistry of peptides, for example, M.Bodanszky, " Peptide Chemistry ", 2nd rev ed., Springer-Verlag, Berlin, Germany, (1993), only be called Bodanszky hereinafter, its content is hereby incorporated by.The example of suitable coupling agent is N, N '-dicyclohexylcarbodiimide, N, I-hydroxybenzotriazole or N-ethyl-N '-[(3-dimethylamino) propyl group] carbodiimide under N '-dicyclohexylcarbodiimide exists.Real and useful coupling agent is commercially available (benzotriazole-1-base oxygen base) three-(dimethylamino) phosphorus hexafluorophosphates, and it exists naturally or exists in I-hydroxybenzotriazole or 4-DMAP.Another kind of reality and useful coupling agent are commercially available 2-(1H-benzotriazole-1-yl)-N, N, N ', N '-tetramethyl-urea a tetrafluoro borate.Another kind of reality and useful coupling agent are commercially available O-(7-azepine benzo triazol-1-yl)-N, N, N ', N '-tetramethyl-urea hexafluorophosphate.

This linked reaction is carried out in inert solvent, for example methylene dichloride, acetonitrile or dimethyl formamide.With excessive tertiary amine, for example diisopropylethylamine, N-methylmorpholine, N-crassitude or 4-DMAP add wherein, are about 8 with the pH value that keeps reaction mixture.Temperature of reaction is usually in 0 ℃～50 ℃ scope and reaction times usually in the scope at 15min～24h.

During linked reaction, must the functional group of component alpha-non-natural amino acid be protected usually, to avoid forming the key of not expecting.Adaptable protecting group is listed in Greene, " ProtectiveGroups in Organic Chemistry ", John Wiley﹠amp; Sons, New York (1981) and " The Peptides:Analysis, Synthesis, Biology ", Vol.3, Academic Press among the New York (1981), only is referred to as Greene hereinafter, and its disclosure is hereby incorporated by.

The α carboxyl of C-terminal residue is usually protected for can cracking providing the ester of carboxylic acid.Operable protecting group comprises 1) alkyl ester; such as methyl, trimethyl silyl and tertiary butyl ester, 2) aralkyl ester, such as benzyl and substituted benzyl ester; perhaps 3) can pass through weak base or gentle method of reducing cracked ester, such as trichloro ethyl ester and phenacyl ester.

Be intended to carry out each amino acid whose alpha-amino group of link coupled and generally carry out N-protected.Can use any protecting group known in the art.Described examples of groups comprises: 1) acyl group, such as formyl radical, trifluoroacetyl group, phthaloyl and ptoluene-sulfonyl; 2) fragrant carbamic acid ester group is such as the bensyloxycarbonyl and the 9-fluorenylmethyloxycarbonyl (Fmoc) of carbobenzoxy-(Cbz) (Cbz or Z) and replacement; 3) aliphatic urethane group is such as tertbutyloxycarbonyl (Boc), ethoxycarbonyl, di-isopropyl methoxycarbonyl and allyloxycarbonyl; 4) cyclic alkyl carbamate group is such as cyclopentyloxy carbonyl and Buddha's warrior attendant alkoxy carbonyl; 5) alkyl is such as trityl and benzyl; 6) trialkylsilkl is such as trimethyl silyl; With 7) contain the group of mercaptan, such as thiophenyl carbonyl and dithio succinyl.Preferred alpha-amino group protecting group is Boc or Fmoc.Many due cares are used for peptide synthetic amino acid derivative and can buy in market.

Generally before next coupling step, described alpha-amino group protecting group is cleaved.When using the Boc group, the method for selection is to use separately or the trifluoroacetic acid in methylene dichloride, perhaps is the HCl in dioxane or ethyl acetate.Then before coupling or original position with basic solution (such as aqueous buffer solution, perhaps the tertiary amine in methylene dichloride or acetonitrile or dimethyl formamide) the gained ammonium salt is neutralized.When using the Fmoc group, the reactant of selection is piperidines or the substituted piperidine in dimethyl formamide, but any secondary amine can use.Described go the protection between 0 ℃～room temperature, carry out, be generally 20～22 ℃.

During using any above-mentioned group to prepare described peptide, generally to protect any natural or alpha-non-natural amino acid with side chain functionalities.The person skilled in the art should be appreciated that for these side chain functionalities, the existence of other blocking group on amino acid and the peptide is depended in the selection of due care base and application.When above-mentioned protecting group is selected, be desirably in during protection and the coupling alpha-amino group, described group is not removed.

For example, when Boc was used as the alpha-amino group protecting group, following Side chain protective group suited: right-tolylsulfonyl part can be used for protection ratio such as Methionin and arginic amino amino side chain; Acetylamino methyl, benzyl (Bn) or tertiary butyl sulphonyl part can be used to protect the sulfide that contains cysteine side chain; Benzyl (Bn) ether can be used to protect the hydroxyl that contains Serine, Threonine or oxyproline side chain; Can be used to protect the carboxyl that contains aspartic acid and L-glutamic acid side chain with benzyl ester.

When selecting Fmoc to be used for α-amine protection, the protecting group based on the tertiary butyl is an acceptable usually.For example, Boc can be used to protect Methionin and arginine, and tertbutyl ether can be used to protect Serine, Threonine and oxyproline and tertiary butyl ester can be used to protect aspartic acid and L-glutamic acid.Trityl (Trityl) part can be used to protect the sulfide that contains cysteine side chain.

Finished in case inhibitor is synthetic in proper order, described any protecting group is just by definite mode is removed by selecting protecting group with any.These methods are the known method of person skilled in the art.

First step in general formula I I compound synthetic generally is functionalized P 1 structural unit of preparation in solution.According to the present invention, can use different compound names.For convenience's sake, use the carbohydrate nomenclature usually at this.The general approach of synthetic two ring P1 groups is by the closed beginning of ring of the intermediate of due care; described intermediate is by 1; 2:5; the different furanose of 6-two-O-isopropylidene-D-obtained by 4 steps; be described in Mayer zum Reckendorf, Chem.Ber.101 (1968) is among the 3802-3807; provided 3S, the stereochemical precursor of 4R.

Scheme 1.a) H ₂, Pd/C, methyl alcohol.B) chloroformic acid benzyl ester, pyridine, methylene dichloride

In scheme 1, the azido-of derivative 1 is reduced into unhindered amina, for example, in the following manner: in suitable solvent, use palladium or other suitable catalyzer on charcoal to reduce by catalytic hydrogenation such as alcohol (for example ethanol or methyl alcohol).The nucleophilic nitrogen-atoms that obtains spontaneous or choose wantonly suitable key (such as, triethylamine or sodium acetate) exist down and the reaction of C-6 position, thereby form 5,5-two encircles.In the entire synthesis process according to compound of the present invention, the leavings group of C-6 is not limited to sulphonate, can also use other leavings group such as halogen.The trinitride residue is reduced into amine can also be undertaken by other method that is known in the document, such as, the usefulness trialkyl-or triaryl phosphorus processing azide derivatives, with the imine derivative of posthydrolysis formation.After the ring closure, amine can carry out N-protected with suitable protecting group, such as carbamate, such as benzyl carbamate or any other similarly common protecting group without acid cleavage of compound 3.Suitable protecting group can be found in: Protective groups in organicchemistry, the third edition, 1999, Theodora W.Greene and Peter G.M.Wuts (Wiley﹠amp; Sons).

For 3R, 4S two ring can use similar method, by can be as TetrahedronAsymmetry, and the 3-azido--3-deoxidation-1 that is prepared described in 10 (1999) 1855-1859,2:5,6-two-O-isopropylidene-D-glucofuranose begins.Then, can described in scheme 2, handle this intermediate.

Scheme 2.a) aqueous acetic acid.B) p-toluenesulfonyl chloride, pyridine, DCM, c) H ₂, Pd/C, methyl alcohol.B) chloroformic acid benzyl ester, pyridine, methylene dichloride.

Compound 4 can be used 5 of selective hydrolysis compound 4, and the weak acid of 6-acetal (such as acetic acid,diluted or similarly acid) is handled, thereby obtains glycol.Primary alconol can selectivity and alkyl-or aryl sulfonyl chloride (such as Tosyl chloride) reaction, thereby provide compound 5.For example, in suitable solvent, use palladium on charcoal or other suitable catalyzer the azido-of derivative 5 to be reduced into unhindered amina by catalytic hydrogenation such as alcohol (for example ethanol or methyl alcohol).The nucleophilic nitrogen that obtains spontaneous or choose wantonly suitable alkali (such as; for example triethylamine or sodium acetate) exist to form with the reaction of C-6 position down and can choose wantonly by 5 of the protecting group N-protected that suits; 5-two ring (such as, its benzyl carbamate (Cbz) for example), thus compound 6 provided.

Additionally, according to scheme 3,3-azido--3-deoxidation-1,2:5,6-two-O-isopropylidene-D-Chinese mugwort Du furanose (idofuranose) (Bull.Chem.Soc.Japan, 57,1 (1984), 237-241) can be 3R, the 4S bicyclic raw material that suits.

Scheme 3.a) aqueous acetic acid.B) p-toluenesulfonyl chloride, pyridine, DCM, c) H ₂, Pd/C, methyl alcohol.B) chloroformic acid benzyl ester, pyridine, methylene dichloride.

Compound 6 can be used 5 of selective hydrolysis compound 6, and the weak acid of 6-acetal (such as acetic acid,diluted or similarly acid) is handled, thereby obtains glycol.Primary alconol can selectivity and alkyl-or aryl sulfonyl chloride (such as Tosyl chloride) reaction, thereby provide compound 7.For example, in suitable solvent, use palladium on charcoal or other suitable catalyzer the azido-of derivative 7 to be reduced into unhindered amina by catalytic hydrogenation such as alcohol (for example ethanol or methyl alcohol).The nucleophilic nitrogen that obtains spontaneous or choose wantonly suitable alkali (such as; for example triethylamine or sodium acetate) exist to form with the reaction of C-6 position down and can choose wantonly by 5 of the protecting group N-protected that suits; 5-two ring (such as, its benzyl carbamate (Cbz) for example), thus compound 8 provided.

Ring is closed to be not limited to the matrix shown in above, can also be used for the derivative shown in the scheme 4.

Scheme 4.a) trinitride is reduced into amine, ring is closed subsequently.B) protection amine.

Rx in the scheme 4 can be selected from methyl, trifluoromethyl, p-methylphenyl or be present in similar residue in the obtainable easily heteroaryl-alkylsulfonyl halides, preferably as Chem.Ber.101 (1968), described in the 3802-3807, be suitable on the primary alconol of glycol, carrying out the large volume Rx of regioselective reaction.R ¹' and R ²' as R ¹And R ²Define.Pg can be the protecting group that suits, such as carbamate (such as benzyl carbamate) or any common similar protecting group without acid cleavage.

Other matrix that is used to encircle closed reaction can be the compound shown in the scheme 5.

Scheme 5.a) trinitride is reduced into amine, ring is closed subsequently.B) protection amine (choosing wantonly).

Rx in the scheme 5 can be selected from methyl, trifluoromethyl, p-methylphenyl or be present in similar residue in the obtainable easily heteroaryl-alkylsulfonyl halides, preferably as Chem.Ber.101 (1968), described in the 3802-3807, be suitable on the primary alconol of glycol, carrying out the large volume Rx of regioselective reaction.R ¹' and R ²' be R as defined above ¹And R ²Ry can be hydrogen or hydroxyl protecting group, the protecting group of preferred ethers type.Preferred Ry is a hydrogen.PG can be the suitable N-protected base of derivative in the scheme 5, and such as carbamate, Ry is generally hydrogen.

Other obtains 5,5-bicyclic method by G.Lin and Z.Shi at Tetrahedron, 53,4,1369-1382, open in 1997.

In scheme 1, obtain 5, other modification of 5-bicyclic compound is shown in the scheme 6.

Scheme 6.a) bromotoluene, sodium hydride, DMF.b)BF ₃.Et ₂O，Et ₃SiH，DCM。c)H ₂，Pd/C，Boc ₂O，1∶1EtOAc-EtOH。D) pyridine, diacetyl oxide.e)H ₂，Pd/C，EtOAc

The acid that compound 9 usefulness are suitable is stablized protecting group (such as the methyl ether that replaces; dibenzyl ether particularly) protects; by with alkali (such as sodium hydride or sodium hydroxide) at aprotic solvent (such as N; dinethylformamide (DMF)) in; the expectation alkylating reagent (such as; benzyl halogenide, particularly bromotoluene) exist the following list-alcohol 9 of handling to obtain.Then, according to G.J.Ewing and M.J.Robins, Org.Lett.1,4,1999, the described method of 635-636 can be reduced into compound 10 with the material that obtains, or pass through reference wherein.Preferably utilize excessive boron trifluoride ethyl ether complex, in the presence of the reductive agent such as trialkyl silane, particularly excessive triethyl silicane reduces in suitable aprotic solvent (such as methylene dichloride).Utilize for example palladium on charcoal, in The suitable solvent or solvent mixture (such as ethyl acetate-ethanol), in hydrogen atmosphere, catalytic hydrogenation compound 10 in the presence of tert-Butyl dicarbonate, in pyridine, handle products obtained therefrom subsequently, provide intermediate 11 with diacetyl oxide.By repeating aforesaid catalytic hydrogenation, obtain list-alcohol 12.

Fluorine can be incorporated on the compound 12 and then according to 7 pairs of bicyclic compounds of scheme carry out N-go the protection.

Scheme 7.a)

Methylene dichloride.B) methyl alcohol sodium methylate.C) 1: 1 methylene dichloride-trifluoroacetic acid.

Compound 13 can use fluorizating agent (such as [two-(2-methoxy ethyl) amino sulphur trifluorides] (

) or (handle such as diethylamino sulphur trifluoride (DAST), by carrying out configuration conversion in the C-5 position, provide product 14 with similar fluorizating agent.Then, by for example handling with methyl alcohol sodium methylate or any similar basic solution with mineral alkali (such as sodium hydroxide or yellow soda ash), compound 14 carries out deacetylated, utilize acidic conditions (such as methylene dichloride-trifluoroacetic acid solution) or other to be found in subsequently: Protective Groups in Organic Chemistry, the third edition, 1999, Theodora W.Greene and Peter G.M.Wuts (Wiley﹠amp; Sons) other method in is carried out the N-deprotection.

In addition, epimeric fluorine can obtain by handle above derivative 9 according to scheme 8.

Scheme 8.a) di-isopropyl azodicarboxylate, phenylformic acid, PPh ₃, THF.B) methyl alcohol sodium methylate.C) bromotoluene, sodium hydride, DMF.D) BF ₃.Et ₂O, Et ₃SiH, methylene dichloride.e)H ₂，Pd/C，Boc ₂O，1∶1EtOAe-EtOH。F) pyridine, diacetyl oxide.g)H ₂，Pd/C，EtOAc。H)

Methylene dichloride.I) methyl alcohol sodium methylate.J) 1: 1 methylene dichloride-trifluoroacetic acid.

Can under the Mitsunobo condition, react accomplished by making compound 16 in the configuration conversion on the C-5, provide benzoic ether.Carry out the ester hydrolysis with the methyl alcohol sodium methylate, handle list-alcohol with bromotoluene subsequently, the epimer 17 of benzyl protection is provided.In the scheme 8 described in reactions steps d-j such as scheme 6 and 7.

Another kind of synthetic wherein R ¹And R ²For the route of " difluoro derivatives " of fluorine is shown in the scheme 9.

Scheme 9.a) bromotoluene, sodium hydride, DMF.B) Et ₃SiH, BF ₃.Et ₂O or trimethylsilyl triflate, DCM.c)H ₂，Pd/C，Boc ₂O，EtOAc-EtOH。D) Benzoyl chloride, pyridine, DCM.e)H ₂，Pd/C，EtOAc。F) Bu ₂SnO, toluene refluxes.G) bromotoluene, cesium fluoride, DMF.H) Dess-Martin crosses iodine alkane.I)

Perhaps diethylamino sulphur trifluoride, DCM.J) methyl alcohol sodium methylate.k)H ₂，Pd/C，EtOAc。1) p-toluenesulfonyl chloride, pyridine, DCM.M) DCM, trifluoroacetic acid.N) triethylamine, methylene dichloride.

The synthetic of P1 structural unit can be by compound 21 (3-azido--3-deoxidation-1, the different furanose of 2-O-isopropylidene-D-) beginning, and described compound 21 is by Mayer zum Reckendorf, Chem.Ber.101 (1968), and 3802-3807 is described.Use benzylating agent, in the presence of alkali, in non-proton property polar solvent (such as N, dinethylformamide), handle compound 21, provide derivative 22 such as sodium hydride or sodium hydroxide such as bromotoluene or benzyl chloride.Then, with trialkyl silane (such as triethyl silicane) and excessive Lewis acid (such as or boron trifluoride ethyl ether complex or trimethylsilyl triflate), in such as the aprotic solvent of methylene dichloride, handle compound 22.Then,, in the presence of tert-Butyl dicarbonate, carry out catalytic hydrogenation, can carry out selective reduction, thereby obtain compound 23 the gained trinitride by utilizing for example palladium on charcoal.In addition, described trinitride can reduce with other method that is known in document, such as triphenylphosphine-water, protects subsequently, provides suitable carbamate.The problem of regioselectivity in the following steps; can use acylating reagent (such as acyl chlorides or acid anhydrides; such as Benzoyl chloride); in pure organic bases (such as pyridine or triethylamine) or mixture (such as dichloride and alkali), compound 23 is handled, thereby provided compound 24 at aprotic solvent.As mentioned above compound 24 is carried out catalytic hydrogenation, provide glycol 25.Selectivity benzylation on the primary alconol of compound 25 can realize by several known in the literature methods.In scheme 9, glycol and dibutyl tin oxide are refluxed in The suitable solvent (such as toluene), thereby form the tin acetal.Then, gained tin acetal can react in DMF with a small amount of excessive bromotoluene and cesium fluoride, provides desired compounds 26.In methylene dichloride, oxygenant (crossing iodine alkane such as the Dess-Martin) oxygenated compound 26 with suitable is converted into secondary alcohol the ketone compound 27 that is suitable for changing into difluoride 28.This step can by with excessive fluorizating agent (such as

) or with diethylamino sulphur trifluoride (DAST) such as methylene dichloride or 1, handle compound 27 in the aprotic solvent of 2-ethylene dichloride and realize.The benzoic ether of compound 28 can carry out cracking with alkali (such as the methyl alcohol sodium methylate), utilizes catalytic hydrogenation to slough benzyl subsequently, thereby obtains glycol 29.Introducing sulphonate in selectivity on the primary alconol can realize by the following method: with a small amount of excessive alkyl-or aryl sulfonyl chloride in the presence of alkali such as pyridine; in such as the suitable solvent of methylene dichloride, handle compound 29; add sulfonylation agent at low temperatures and slowly temperature is increased to room temperature, provide list-alcohol 30.Under the acidic conditions of methylene dichloride-trifluoroacetic acid, handle compound 30 and discharge amine and use this product of alkaline purification to promote the inner loop closure, provide structural unit 31 such as triethylamine.

In addition synthetic 5,5-bicyclic route is shown in scheme 10 and 11.

Scheme 10.a) fluorizating agent.B) reduction amine or N-deprotection, the optional N-protected that carries out subsequently.C) reductive agent.

In scheme 10; such as the substituting group in C-3 and C-4 position is the derivative of the compound 32 (can obtain or utilize method well known in the art to obtain as mentioned above) of cis relation; Lg is the leavings group such as halogen or sulphonate; equal trinitride with R or with the nitrogen of suitable N-protected base protection; can form compound 33 with handling such as above-mentioned fluorizating agent.By or reduction trinitride or discharge the amine of being sheltered by suitable deprotection method, described amine can carry out the intramolecularly attack on the C-6 position, produce 5 of structure 34,5-two rings, it can be chosen wantonly and carry out N-protected (Pg=protecting group or hydrogen).Use such as above-mentioned suitable reductive agent or with similar reductive agent reduction C-1 and will provide structural unit 35.

In scheme 11, another kind of synthetic two fluoro-5 have been shown, 5-bicyclic route.

Scheme 11.a) oxidation.B) fluorizating agent.C) reduction trinitride or N-deprotection, the optional N-protected that carries out subsequently.D) reductive agent.

In scheme 11; substituting group in C-3 and C-4 position is the compound 36 (can obtain or utilize method well known in the art to obtain as mentioned above) of cis relation; Lg is the leavings group such as halogen or sulphonate; equal trinitride with R or with the nitrogen of suitable N-protected base protection; can utilize Swern type reaction or other suitable method to carry out oxidation, thereby can provide compound 37.According to scheme 11, handle compound 37 with excessive such as above-mentioned fluorizating agent, provide compound 38.Discharge 38 the amine of being sheltered by the reduction trinitride or by suitable deprotection method, described amine can carry out the intramolecularly attack on C-6, forms 5 of structure 39,5-two rings, and it can be chosen wantonly and carry out N-protected (Pg=protecting group or hydrogen).Use such as above-mentioned suitable reductive agent or with similar reductive agent reduction C-1 and will provide structural unit 40.

Synthetic wherein R ¹Perhaps R ²For the suitable route of the compound of halogen (such as chlorine) is shown in the scheme 12.

Scheme 12

A) thionyl chloride, b) methyl alcohol sodium methylate, c) 1: 1 methylene dichloride-trifluoroacetic acid, d) thionyl chloride, pyridine.

The P1 structural unit is generally by conventional liquid phase or solid state chemistry process, such as disclosed chemical process among the chemical process of following discloses or illustration or WO00/69855 or the WO02/057270, prolong (perhaps P3+P2 structural unit) with natural or non-natural P2 amino acid.P2 and P3 group or commercially available for enantiomer or can or can utilize the known simple chemovar of those skilled in the art to obtain from racemate resolution.For example, 4-(methyl-piperazine-1-yl)-phenylformic acid can utilize Buchwald chemical process (S.L.Buchwald﹠amp; J.P.Wolfe, Journal of Organic Chemistry, 2000,65,1144) obtain and carry out precision work subsequently.Other P3 nuclear utilizes the Friedel-Crafts acylation reaction to be prepared by 1-(4-phenyl-piperidines-1-yl)-ethyl ketone and utilizes the known standard chemical modification of those skilled in the art to carry out precision work subsequently such as 4-(1-piperidin-4-yl)-phenylformic acid.Additionally, other P3 part is such as 5-[2-(4-morpholinyl) oxyethyl group]-2-cumarone-2-carboxylic acid utilizes the Mitsunobu reaction that solid phase carries out to be prepared, as L.S.Richter﹠amp; T.R.Gadek is at Tetrahedron Lett., in 1994,35,4705 describe in detail.

The typical case of scheme 13. cyclic ketones prolongs

Additionally, for the P1 structural unit of hydroxyl can prolong and carries out oxidation subsequently, shown in scheme 14.

Scheme 14, the typical case of hydroxylated P1 structural unit prolongs.

The P3 end is general by making the midbody compound of following formula

R wherein ⁶With Rc as defined above and LG be conventional leavings group, such as triflate or the like, prolong with the P1/P2 structural unit reaction of N-deprotection shown in above.This is reflected in the suitable organic solvent and carries out, include but not limited to that halogenated organic solvent is (such as methylene dichloride and 1,2-ethylene dibromide or the like), ether solvent (such as ether, tetrahydrofuran (THF)), acetonitrile or aromatic solvent (such as benzene, toluene and dimethylbenzene or the like) or its mixture and choose wantonly in the presence of organic or mineral alkali and carry out.Preferred organic bases is triethylamine, pyridine, N-methylmorpholine, picoline and diisopropylethylamine or the like.Preferred described mineral alkali is cesium carbonate, yellow soda ash and sodium bicarbonate or the like.This reaction is chosen wantonly in the presence of the siccative such as molecular sieve and is carried out.This reaction is preferably at room temperature carried out.Intermediate can be prepared by method well known in the art.For example, R wherein ⁶Be phenyl or 4-fluorophenyl, Rb is that trifluoromethyl and Rc are that the compound of hydrogen can be respectively by commercially available 2,2,2-trifluoroacetophenone or 2,2,2,4 '-the tetra fluoro benzene ethyl ketone is prepared easily, by suitable reductive agent (such as sodium borohydride and lithium aluminum hydride or the like) ketone group is reduced into alcohol radical.The solvent that uses depends on the type of reductive agent.For example, when using sodium borohydride, be reflected in the alcohol organic solvent and carry out, such as methyl alcohol and ethanol or the like.For example, when using lithium aluminum hydride, be reflected in the ether solvent and carry out, such as tetrahydrofuran (THF) or the like.2,2,2-three fluoro-1-phenylethyl alcohols or 2,2, the reaction of 2-three fluoro-1-(4-fluorophenyl) ethanol and trifluoromethanesulfanhydride anhydride provides desired compounds.The intermediate of chirality enrichment can obtain in the following manner: suitable catalyzer (such as (and A or (R) the CBS catalyzer or (A or (R)-, a-phenylbenzene-2-tetramethyleneimine-methyl alcohol) exists down, in the presence of BBN, use suitable reductive agent (such as catecholborane or BH ₃-DMS title complex) the corresponding benzene halide ethyl ketone of reduction.

In corresponding mode, the following formula intermediate:

Can react with the P2 structural unit of carboxy protective, it can carry out deprotection subsequently and prolong with P1 structural unit described herein.

Additionally, above-mentioned intermediate:

LG is suitable leavings group, and the hydroxyl protecting group for suiting such as triflate and PG such as trialkylsilkl or the like, reacts under above-mentioned reaction conditions.The O-protection hydroxyl ethanamide of gained is oxidized to corresponding carboxylic acid and is coupled on the P1 structural unit as described below.Suitable hydroxyl protecting group and be used to make them to protect and the reaction conditions that they are removed can be found in Greene, T.W.; And Wuts, P.G.M.Protecting Groups in OrganicSynthesis; John Wiley﹠amp; Sons is among the Inc.I 999.The P2 oxyethylamine can be prepared by method well known in the art by corresponding natural or alpha-non-natural amino acid.Some described methods are described among the open No.WO 03/075836 of PCT application, and its whole disclosures are hereby incorporated by.

Additionally, wherein E can be by making following formula: compound for the compound of-CRbRc-

R wherein ⁶For cyclic group and Rb as defined above are halogenated methyl, preferred trifluoromethyl reacts under the reductive amination reaction conditions with the P2 structural unit of N-deprotection, carboxyl-protection or P1/P2 structural unit as implied above.This is reflected at suitable dewatering agent (such as TiCl4, sal epsom, sec.-propyl trifluoroacetate) existence down, is carrying out in the presence of the alkali (such as diisopropylethylamine and pyridine or the like) and in suitable organic solvent (such as methylene dichloride), thereby is providing imines.Imines reduces in suitable organic solvent (such as methyl alcohol and ethanol or the like) with suitable reductive agent (such as sodium borohydride and sodium cyanoborohydride or the like).

Additionally, wherein E can be prepared by haloalkyl aldehyde and amine as follows are reacted for the compound of-CRbRc-:

R ⁵, R ⁵', R6 and Rb as defined above.Use the Dean-Stark device, make haloalkyl aldehyde and monoethanolamine (being prepared) condensation by under condition well known in the art, reduce corresponding 5/5 ' alpha amino acid with suitable reductive agent (such as lithium aluminum hydride or the like), cyclic acetal amine shown in providing, its by with formula R ⁶Grignard reagent or the formula R of MgX (wherein X is a halogen) ⁶The organolithium reagent reaction of LiI, the hydroxyl acetamide shown in providing.With suitable oxygenant (such as Jones oxygenant or H ₅IO ₆/ CrO ₃Or the like) the oxidation hydroxyl acetamide, the P3/P2 structural unit is provided then, its C-is terminal to prolong and carries out as required oxidation with the P1 structural unit.

Aforesaid prolongation is generally in the presence of suitable coupling agent, benzotriazole-1-base oxygen base tripyrrole alkane phosphorus hexafluorophosphate (PyBOP) for example, O-benzotriazole-1-base-N, N, N ', N '-tetramethyl--urea hexafluorophosphate (HBTU), O-(7-azepine benzo triazol-1-yl)-1,1,3,3-tetramethyl--urea hexafluorophosphate (HATU), 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDC) or 1,3-dicyclohexylcarbodiimide (DCC), choose wantonly I-hydroxybenzotriazole (HOBT) and alkali (such as N, the N-diisopropylethylamine, triethylamine, N-methylmorpholine or the like) carries out under the existence.This reaction is generally carried out under 20～30 ℃, preferably carries out under about 25 ℃, and needs 2～24 hours to finishing.Suitable reaction solvent is an inert organic solvents, such as halogenated organic solvent (for example, methylene dichloride and chloroform or the like), acetonitrile, N, and dinethylformamide, ether solvent (such as tetrahydrofuran (THF) with diox or the like).

Additionally, above-mentioned prolongation coupling step can carry out in the following manner: at first the P3/P2 structural unit is changed into active acid derivant, such as succinimide ester, itself and P1 amine are reacted.This reaction generally needed react completely in 2～3 hours.The condition of using in this reaction depends on the person's character of active acid derivant.For example, if it is a chloride derivative 4, this is reflected under suitable alkali (for example triethylamine, diisopropylethylamine and pyridine or the like) existence and carries out.Suitable reaction solvent is a polar organic solvent, such as acetonitrile, N, and dinethylformamide, methylene dichloride or its any suitable mixture.

Utilize aforesaid technology, aforesaid method can also be used to prepare wherein that Rc is not the compound of hydrogen, but uses formula R ⁶The ketone of RbCO is replaced R ⁶COH and handle the cyclic acetal amine of gained then with RcLi/RcMgX carries out oxidation subsequently, thereby provides free acid.Then, make free acid under aforesaid condition, carry out condensation.

To those skilled in the art, obviously wherein E be that the compound of CRbRc can also be prepared as follows:

Particularly, conventional O-protects above-mentioned monoethanolamine, and the haloalkyl group with imine moiety shown in providing with the reaction of haloalkyl hemiacetal is subsequently used R ⁶The organolithium compound of Li is handled it, wherein R ⁶As defined above.Remove the deoxidation protecting group and be provided at the hydroxyl acetamide described in the scheme, in the corresponding way it is oxidized to carboxylic acid and prolongs with the P1 structural unit.Suitable oxygen protecting group and be used to make them to protect and the reaction conditions that they are removed can be found in Greene, T.W.; And Wuts, P.G.M.Protecting Groups in OrganicSynthesis; John Wiley﹠amp; Sons is among the Inc.I 999.

Additionally, wherein E is CRbRc and R ⁶For the compound of aryl or heteroaryl can as followsly be prepared:

Particularly, make the reaction of above-mentioned haloalkyl hemiacetal and shielded P2 structural unit, thus the 2-shown in obtaining (1-methylol amino) acetic ester intermediate.This acid (such as tosic acid) that is reflected at catalytic amount exists down and carries out in aromatic hydrocarbon solvent (such as toluene and benzene or the like).

At Friedel-Crafts reaction conditions/BF ₃.EtT ₂Under the O, use R ⁶H handles 2-(1-methylol amino) acetic ester intermediate the P3/P2 structural unit of carboxy protective is provided, and it is prolonged.Similarly, can make haloalkyl hemiacetal and P2/P1 structural unit the reaction and as required it is oxidized to ketone.

Term " N-protected base " or " N-protected " are meant that those N-that are intended to protect amino acid or peptide are terminal or protection is amino as used herein, make it that group of the reaction do not expected not take place during synthesis technique.Usually the N-protected base of using is disclosed in Greene, " ProtectiveGroups in Organic Synthesis " (John Wiley﹠amp; Sons, New York, 1981) in, it is hereby incorporated by.The N-protected base comprises acyl group, such as formyl radical, ethanoyl, propionyl, pivalyl, tertiary butyl ethanoyl, 2-chloracetyl, 2-acetyl bromide, trifluoroacetyl group, tribromo-acetyl base, phthaloyl, neighbour-nitro-phenoxy ethanoyl, α-chlorobutyryl, benzoyl, 4-chlorobenzene formacyl, 4-benzoyl bromide and 4-nitro benzoyl or the like; Alkylsulfonyl is such as benzenesulfonyl and ptoluene-sulfonyl or the like; Carbamate forms group, such as carbobenzoxy-(Cbz), right-the benzyloxycarbonylchloride base, right-methoxyl group benzyloxy carbonyl, right-the nitro carbobenzoxy-(Cbz), 2-nitro carbobenzoxy-(Cbz), right-bromo-benzyloxycarbonyl, 3, the 4-dimethoxy-benzyloxycarbonyl, 4-methoxyl group benzyloxy carbonyl, 2-nitro-4, the 5-dimethoxy-benzyloxycarbonyl, 3,4,5-trimethoxy carbobenzoxy-(Cbz), 1-(right-xenyl)-1-methyl ethoxy carbonyl, α, alpha-alpha-dimethyl-3, the 5-dimethoxy-benzyloxycarbonyl, the hexichol methoxycarbonyl, tertbutyloxycarbonyl, the di-isopropyl methoxycarbonyl, the different third oxygen carbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2, the 2-trichloro-ethoxycarbonyl, carbobenzoxy, 4-nitro carbobenzoxy, fluorenyl-9-methoxycarbonyl, the cyclopentyloxy carbonyl, adamantyl oxygen base carbonyl, cyclohexyl oxygen base carbonyl and thiophenyl carbonyl or the like; Alkyl is such as benzyl, trityl and benzyloxymethyl or the like; And silyl, such as trimethyl silyl or the like.Preferred N-protected base comprises formyl radical, ethanoyl, benzoyl, pivalyl, uncle's butyryl radicals, benzenesulfonyl, benzyl, tertbutyloxycarbonyl (BOC) and carbobenzoxy-(Cbz) (Cbz).

Hydroxyl and/or carboxyl-protecting group have also carried out extensive overview in the same Greene; and comprise ether; such as methyl; substituent methyl ether; such as methoxymethyl; methylthiomethyl; benzyloxymethyl; tert.-butoxy methyl and 2-methoxy ethoxy methyl or the like; silyl ether; such as trimethyl silyl (TMS); t-butyldimethylsilyl (TBDMS); the tribenzyl silyl; triphenylsilyl; t-butyldiphenylsilyl and triisopropyl silyl or the like; the ether that replaces; such as the 1-ethoxyl methyl; 1-methyl isophthalic acid-methoxy ethyl; the tertiary butyl; allyl group; benzyl; to methoxy-benzyl; diphenyl methyl and trityl or the like; aralkyl; such as trityl; and pixyl (9-hydroxyl-9-phenyl Xanthene derivative, particularly muriate).The ester hydroxyl protecting group comprises ester, such as manthanoate, benzyl manthanoate, chloracetate, methoxyacetic acid ester, phenylium ester, pivalate, diamantane ester,

Acid esters and benzoic ether or the like.The carbonic ether hydroxyl protecting group comprises methyl ethylene, allyl group, cinnamyl and benzyl or the like.

The detailed description of embodiment

With reference now to following examples,, only the mode by illustration is described multiple embodiments of the present invention.

Embodiment 1

The structure of P1 structural unit

Step a)

Under slight positive pressure, to 54 (5.2g, 13.0mmol), carbon carry palladium (10%, Acros, carbinol mixture 0.66g) carries out hydrogenation.In 1 hour time, hydrogen is changed 3 times, show that at TLC (petroleum ether-ethyl acetate 7: 3 and methylene chloride-methanol 9: 1, painted) raw material changes into the active spot of main non-UV and some more weak higher mobile spots (methylene chloride-methanol 9: 1) of painted AMC fully with ammonium molybdate-cerous sulfate.Then, with the reaction mixture filtration over celite and concentrate, provide crude compound 55.

Under 0 ℃, to the methylene dichloride (60ml) of gained resistates and pyridine (3.2ml, add in suspension 40mmol) chloroformic acid benzyl ester (0.93ml, 6.5mmol).At room temperature reaction mixture was stirred 2 hours, after this, down other pyridine (3ml) and chloroformic acid benzyl ester (0.8ml) are added wherein at 0 ℃.Then, at room temperature above-mentioned reaction mixture is stirred and spend the night, use methylene dichloride (100ml) dilution then, order with the 1M aqueous sulfuric acid (2 * 50ml) and the 1M sodium bicarbonate aqueous solution (1 * 50ml) washing is carried out drying (sodium sulfate) then, filters and is concentrated on silicon-dioxide.Use ethyl acetate: 1: 4 (350ml), 2: 3 (250ml) of sherwood oil, 1: 1 (250ml), 3: 2 (250ml) carry out flash chromatography with 3: 1 (150ml) to the gained resistates and separate (diameter: 4cm, YMC-gel: 50g, fill elutriant: ethyl acetate: sherwood oil 1: 4), in a vacuum after the dried overnight, be given the compound 56 (2.71g of spumescence slurries, 8.1mmol, 2 step productive rates 62%).

NMR data (400MHz, CDCl ₃): 1H, 1.33,1.52 (2s, 6H, C (CH ₃) ₂), 2.34 (2d, 1H ,-OH), 3.04 (m, 1H, H-6a), 3.97 (m, 1H, H-6b), 4.19 (m, 1H, H-5), 4.33 (m, 1H, H-3), 4.68,4.84 (2d, 1H, H-2), 4.79 (t, 1H, H-4), 5.08-5.24 (m, 2H, CH ₂Ph), 5.86 (br s, 1H, H-1), 7.30-7.42 (m, 5H, Ar-H).

Step b)

In 5 fens clock times, to the sodium hydride that stirs (be 60% in mineral oil, Aldrich, 0.34g, 8.4mmol) and compound 56 (2.17g, add in dimethyl formamide 6.47mmol) (30ml) suspension bromotoluene (0.81mmol, 6.8mmol).Stir after 1 hour (TLC: ethyl acetate: sherwood oil 2: 3), methyl alcohol (approximately 2ml) is added wherein to destroy excess reagent, immediately it is distributed between ethyl acetate (180ml) and the water (150ml) then.Gained organic layer water (carry out drying (sodium sulfate) then, filter and concentrate on silicon-dioxide by 3 * 100ml) washings.Use ethyl acetate: 1: 4 (100ml) of sherwood oil, 3: 7 (250ml) carry out flash chromatography with 2: 3 (250ml) to resistates and separate (diameter: 4cm, YMC-gel: 40g, fill elutriant: ethyl acetate: sherwood oil 1: 4), in a vacuum after the dried overnight, provide colourless slurries (2.7g, 6.35mmol, 98%).

NMR data (400MHz, CDCl ₃): 1H, 1.31 (s, 3H, C (CH ₃) (CH ₃)), 1.51 (d, 3H, C (CH ₃) (CH ₃)), 3.29 (m, 1H, H-6a), 3.78-3.96 (m, 2H, H-5 and H-6b), 4.22 (dd, 1H, H-3), 4.64,4.84 (2M, 4H, H-2, H-4 and CH ₂Ph), 5.07-5.22 (m, 1H, CH ₂Ph), 5.94 (m, 1H, H-1), 7.28-7.39 (m, 10H, Ar-H).

Step c)

Under 0 ℃, to the compound 7 that stirs (2.635g, methylene dichloride 6.19mmol) (28ml) and triethyl silicane (9.9ml, 61.9mmol) once add in the solution boron trifluoride diethyl etherate title complex (7.9ml, 61.9mmol).Then, at room temperature above-mentioned reaction mixture is stirred 24h (TLC: petroleum ether-ethyl acetate 4: 1 and ethyl acetate-toluene 3: 2), then 1M sodium bicarbonate aqueous solution (40ml) and some solid sodium bicarbonates are added wherein carefully, till stopping bubbling.The gained mixture is distributed between methylene dichloride (100mL) and the water (100mL).The gained organic layer with the 1M sodium bicarbonate aqueous solution (1 * 100ml) and salt solution (1 * 100ml) washing is carried out drying (sodium sulfate), filtration then and is concentrated on silicon-dioxide.Use ethyl acetate: toluene 3: 2 (750ml), carry out flash chromatography to resistates and separate (diameter: 4cm, YMC-gel: 48g, fill elutriant: ethyl acetate-toluene 3: 2),, provide the colourless hard stock liquid (1.38g of about 85-90% purity according to TLC, 3.74mmol, 60%).LR-MS:C ₂₁H ₂₄NO ₅: calculated value 370.2. measured value: 370.0[M+H].

Step d)

At slight excessive rolling, to compound 58 (1.38g, 3.74mmol), carbon carry palladium (Acros, 10%, 0.12g) and tert-Butyl dicarbonate (0.82g, ethyl acetate 3.7mmol) (50ml) mixture carries out hydrogenation.In 1 hour time, hydrogen is changed twice, and reaction is monitored by LC-MS.After 1 hour, other carbon is carried palladium (0.1g) add wherein, and reaction mixture is in addition with hydrogen treat 1 hour again.Then, with the reaction mixture filtration over celite and concentrate.The gained resistates spends the night with the processing of 2: 1 pyridine-diacetyl oxides (18ml), concentrates then.The gained resistates is dissolved in the methylene dichloride (60ml) again, and with the 1M aqueous sulfuric acid (2 * 40ml) and the 1M sodium bicarbonate aqueous solution (1 * 40ml) order washing is carried out drying (sodium sulfate) then, filters and is concentrated.Use ethyl acetate: toluene 1: 4 (200ml) and 1: 3 (150ml) carry out flash chromatography to resistates (be dissolved in toluene-ethyl acetate 4: 1 in) and separate (diameter: 3cm, YMC-gel: 20g, fill elutriant: ethyl acetate: toluene 1: 4), in a vacuum after the dried overnight, provide colourless slurries (1.13g, 3.0mmol, 80%).

NMR data (400MHz, CDCl ₃): 1H, 1.45 (s, 9H, C (CH ₃) ₃), 2.08 (s, 3H, COCH ₃), 3.10 (m, 1H, H-6a), 3.74-3.99 (m, 3H, H-1a, H-5 and H-6b), 4.11 (m, 1H, H-1b), 4.16-4.74 (m, 4H H-3, H-4 and CH ₂Ph), 5.31 (m, 1H, H-2), 7.28-7.40 (m, 5H, Ar-H).

Step e)

At slight excessive rolling, with compound 60 (1.08g, 2.86mmol) and carbon carry palladium (10%, ethyl acetate 0.15g) (30ml) mixture hydrogenation 2h (TLC: toluene-ethyl acetate 4: 1 and 1: 1), then with its filtration over celite and concentrate.(in 3 * 10ml) mixture is concentrated, then it is dissolved in the methylene dichloride and under 0 ℃, in this solution, add two-(2-methoxy ethyl) amino sulphur trifluorides (being 50%, 2.12ml, 2 equivalents) in THF at methylene dichloride.After at room temperature stirring is spent the night, with the other amino sulphur trifluoride of two (2-methoxy ethyls) (is 50% in THF, 2ml) (TLC: toluene-ethyl acetate 1: 1 wherein and is at room temperature stirred reaction mixture one night in adding in addition, triketohydrindene hydrate is painted), then the 1M sodium bicarbonate aqueous solution is added wherein carefully, till stopping bubbling.The gained mixture washs once with 1M sodium bicarbonate aqueous solution (40ml) with methylene dichloride (50ml) dilution and gained organic layer, carries out drying (sodium sulfate) then, filters and concentrates.Use 4: 1 pairs of gained resistatess of toluene-ethyl acetate (be dissolved in toluene-ethyl acetate 4: 1 in) to carry out flash chromatography and separate (diameter: 3cm, silicon-dioxide: 25g, fill elutriant: toluene-ethyl acetate 4: 1), in a vacuum after the dried overnight, be given the compound 62 (0.49g of colourless slurries, 1.7mmol, 59%).Some raw materials and sulphur intermediate can reclaim from reaction mixture.

LR-MS:C ₉H ₁₃FNO ₅: calculated value 234.1. measured value: the 234.0[M+2H-tertiary butyl].

Embodiment 2

Prolong with typical P2

Step a)

(0.49g adds 0.5M methyl alcohol sodium methylate (1ml) in methyl alcohol 1.7mmol) (9.5ml) solution, at room temperature it is stirred 30 minutes (TLC: toluene-ethyl acetate 3: 2, triketohydrindene hydrate is painted) then to compound 62.Dowex W X 8 (50-100 mesh, H with methanol wash ⁺-form) adds wherein (the pH value is monitored by the pH-test paper) carefully, till reaching neutrality, then mixture is filtered and concentrates.Resistates is dissolved in the methylene dichloride, and under 0 ℃, trifluoroacetic acid is added wherein.At room temperature reaction mixture is stirred 55 minutes (TLC: methylene chloride-methanol 9: 1, triketohydrindene hydrate is painted), concentrate then.Use methyl alcohol: 5: 95 (150ml) of methylene dichloride, 7: 93 (100ml) and 1: 9 (200ml), resistates (be dissolved in methylene chloride-methanol 95: 5 in) is carried out column chromatography separate (diameter: 2cm, silicon-dioxide: 15g, fill elutriant: methylene chloride-methanol 95: 5), by placing crystallization, provide hard stock liquid (0.39g, 1.50mmol, 88%).

NMR data (400MHz, DMSO-d ₆): 1H, 3.34,3.44 (2dd, 1H, H-6a), 3.60-3.70 (m, 2H, H-1a and H-6b), 3.89 (dd, 1H, H-1b), 4.15 (d, 1H, H-3), 4.51 (br s, 1H, H-2), 4.76 (dd, 1H, H-4), 5.26 (dd, ²J _{H, F}=48.3Hz, H-5).

Step b)

To the compound 64 (0.34g that stir, 1.30mmol), N-ethyl-N '-(3-dimethylaminopropyl) carbodiimide hydrochloride (0.28g, 1.43mmol), I-hydroxybenzotriazole hydrate (0.22g) and N-(tertbutyloxycarbonyl)-L-leucine monohydrate (0.34g, 1.37mmol) DMF (10ml) solution in add triethylamine (0.54ml, 3.9mmol), at room temperature it was stirred 24 hours then.Above-mentioned reaction mixture is distributed between 10% aqueous citric acid solution (30ml) and the ethyl acetate (10ml).The gained water layer with ethyl acetate (3 * 10ml) extract, then organic layer is merged and the order water (1 * 20ml) and the 1M sodium bicarbonate aqueous solution (3 * 20ml) washings are carried out drying (sodium sulfate) then, filter and are concentrated on silicon-dioxide.Utilize the petroleum ether solution (40-60%, substep gradient elution) of ethyl acetate that the gained resistates is carried out the flash chromatography separation, be given 15 (0.35g, 0.98mmol, 75%) of colourless amorphous solid.

LR-MS:C ₁₃H ₂₂FN ₂O ₅: calculated value 305.1. measured value: the 305.1[M+2H-tertiary butyl].

Embodiment 3

Be oxidized to P1 ketone.

This embodiment has shown the oxidation of the hydroxylated P1 compound of model P3+P2+.

At room temperature, to the compound 66 that stirs (0.10g, add in methylene dichloride 0.25mmol) (4ml) solution Dess-Martin cross iodine alkane (0.12g, 0.28mmol).After stirring 90 minutes, the gained reaction mixture dilutes with methylene dichloride (10ml), with 1: 11M sodium bicarbonate aqueous solution-10% sodium thiosulfate solution (carry out drying (sodium sulfate) then, filter and concentrate on silicon-dioxide by 4 * 10ml) washings.Utilize the petroleum ether solution (50-60%, substep gradient elution) of ethyl acetate that the gained resistates is carried out the flash chromatography separation, be given 67 (0.072g, 0.18mmol, 71%) of colourless foam.Compound 67 is obtained to be the mixture of geometrical isomer (rotational isomer) and their hydrate.

LR-MS:C ₂₁H ₂₄FN ₂O ₅: calculated value 403.2. measured value: 403.0[M+H] the keto-acid NMR sample of .67 obtains as follows; (geometrical isomer and hydrate forms ratio are: 6: 4 mixture of hydrate/ketone) be dissolved among the DMSO-d6 with 5mg compound 67, in the NMR device, be heated to 100 ℃ then, make it reach 50 ℃ then, take this NMR and only represent that the hydrate forms of trace and the ratio of rotational isomer are 2: 1.

NMR data (500MHz, DMSO-d6,50 ℃): ¹H, 0.90-1.04 (m, 4x CH ₃, main and less important form), 1.39-1.82 (m, 2x CH ₂CH (CH ₃) ₂With 2x CH ₂CH (CH ₃) ₂, main and less important form), 3.56 (m, H-6a, less important), 3.82 (m, H-6A, main), 3.97-4.25 (m, 4x H-1, main and less important form and H-6b, less important), 4.37 (dd, H-6b, main), 4.62 (d, H-3, less important), 4.79 (m, H, main), 4.84 (d, H-3, main), 4.94 (m, H-4, main), 5.12 (m, H-4, less important), and 5.15-5.34 (m, H-5 is main and H-5 is less important, and H is less important, J _{H, F is main}=49.1Hz, J _{H, F is less important}=49.4Hz), 7.35 (t, 1H, Ar-H), 7.47 (t, 1H, Ar-H), 7.57-7.70 (m, 2H, Ar-H), 7.78 (d, 1H, Ar-H), 8.18 (d ,-NH, less important), 8.70 (d ,-NH, main).

Embodiment 4

Alternative P1 epimer.

Step a)

At room temperature, (1.58g, the methanol solution (5mL) of adding 0.5M sodium methylate stirs them 40 minutes then in methyl alcohol 4.19mmol) (20mL) solution to the compound (60) that stirs.Then, reaction mixture Dowex 50WX 8 (H ⁺-form) neutralization is filtered, and adds triethylamine to slight alkalescence, concentrates then and (concentrates in 2 * 20mL) at toluene.Under 0 ℃, (0.43g, (0.76g 5.02mmol), at room temperature spends the night its stirring then to add TERT-BUTYL DIMETHYL CHLORO SILANE in DMF 6.28mmol) (10mL) solution to the resistates that stirs and imidazoles.Then, reaction mixture dilutes with ethyl acetate (100mL), the order with 10% aqueous citric acid solution (3 * 50mL) and the 1M sodium bicarbonate aqueous solution (3 * 50mL) washing, carry out drying (sodium sulfate), the filtration and on silicon-dioxide, concentrate.The gained resistates is carried out the column chromatography separation, and (substep gradient elution, the toluene solution of ethyl acetate 5-20%), are provided as the intermediate (1.86g) of protection fully of slurries.Palladium that will be on charcoal (Aldrich 10%, 0.28g) and the intermediate of above acquisition (1.80g, ethyl acetate 4.00mmol) (40mL) mixture be slight excessive rolling hydrogenation 1 hour, then with its filtration over celite and concentrate.

By drying the gained material is carried out crystallization in a vacuum, be provided as 72 (1.34g, 90%) of aciculiform thing.

NMR data (400MHz, CDCl ₃): 1H, δ 0.14 (m, 6H, Si (CH ₃) ₂), 0.90 (m, 9H, SiC (CH ₃) ₃), 1.48 (m, 9H, C (CH ₃) ₃), 2.53 (m, 1H, OH), 2.78 (dd, 1H ,-H-6A), and 3.67-4.05 (m, 3H, H-1A, H-1B and H-6B), 4.05-4.21 (m, 2H, H-3 and H-5), 4.35-4.50 (2brs, 1H, H-2), 4.57 (m, 1H, H-4).

Step b)

Under 0 ℃, in 20 fens clock times, to (the 72) (1.068g that stirs, 2.97mmol), phenylformic acid (0.50g, 4.46mmol) and triphenylphosphine (1.17g, 4.46mmol) THF (15mL) solution in drip to add di-isopropyl azodicarboxylate (0.88mL, THF 4.46mmol) (5mL) solution.Then, at room temperature above-mentioned reaction mixture is stirred and spend the night, on silicon-dioxide, concentrate then.Use petroleum ether-ethyl acetate as elutriant gained resistates to be carried out flash chromatography at 9: 1 and separate, provide colourless slurries (1.34g, 97%).

NMR data (400MHz, CDCl ₃): 1H, δ 0.08-0.21 (m, 6H, Si (CH ₃) ₂), 0.90 (s, 9H, SiC (CH ₃) ₃), 1.42-1.56 (m, 9H, C (CH ₃) ₃), 3.48 (m, 1H, H-6A), 3.70-4.01 (m, 3H, H-1A, H-1B, H-6B is less important and main), 4.21,4.30 (2d, 1H, H-3), 4.44,4.56 (2brs, 1H, H-2), 4.72 (m, 1H, H-4), 5.34 (d, 1H, H-5), 7.45 (t, 2H, Ar-H), 7.58 (t, 1H, Ar-H), 8.00 (d, 2H, Ar-H).

Step c)

At room temperature, (1.34g, the methanol solution (6mL) of adding 0.5M sodium methylate stirs them 15 minutes then in methyl alcohol 2.89mmol) (6mL) solution to the compound 73 that stirs.Then, reaction mixture Dowex 50WX 8 (H ⁺-form) neutralizes and filter.The solution that obtains joined be similar to that above (0.187g 0.40mmol) in the solution that begins to obtain, concentrates then by (II).Use toluene-ethyl acetate as elutriant gained resistates to be carried out flash chromatography at 3: 2 and separate, carry out crystallization, be given 74 (1.091g, 92%) of colourless slurries by drying in a vacuum.

NMR data (400MHz, CDCl ₃): 1H, δ 0.06-0.20 (m, 6H, Si (CH ₃) ₂), 0.89 (s, 9H, SiC (CH ₃) ₃), 1.42-1.54 (m, 9H, C (CH ₃) ₃), 2.03 (brs, 1H, OH), 3.28 (dd, 1H, H-6A), 3.53-3.79 (m, 3H, H-1A, H-1B, H-6B), 4.19 and 4.34-4.56 (2m, 4H, H-2, H-3, H-4 and H-5).

Step d)

At room temperature, (0.428g adds Deoxofluor and (is 50% in THF, 0.53mL), causes slight intensification in methylene dichloride 1.19mmol) (10mL) solution to (74) of stirring in the flask of Teflon coating.At room temperature the gained reaction mixture was stirred 72 hours, use methylene dichloride (20mL) dilution then, (2 * 20mL) washings are carried out drying (sodium sulfate), filter and are concentrated on silicon-dioxide with the 1M sodium bicarbonate aqueous solution.Use petroleum ether-ethyl acetate as elutriant gained resistates to be carried out flash chromatography at 9: 1 and separate, be given (IV) (0.118g, 27%) of water white oil.

NMR data (400MHz, CDCl ₃): 1H, δ 0.08-0.20 (m, 6H, Si (CH ₃) ₂), 0.89 (s, 9H, SiC (CH ₃) ₃), 1.42-1.53 (m, 9H, C (CH ₃) ₃), 3.26 and 3.36 (2dd, 1H, H-6A), 3.64 (m, 1H, H-1A), 3.73-4.04 (m, 3H, H-1B, H-6B), 4.20 (dd, 1H, H-3 ^*), 4.40,4.51 (2s, 1H, H-2), 4.69 (m, 1H, H-4 ^*) 4.86,4.98 (2brs, 1H, H-5). ^*Can be exchanged.

Step e)

(0.229g, the THF solution (0.70mL) of adding 1M tetrabutyl ammonium fluoride at room temperature stirs them 40 minutes then in THF 0.63mmol) (8mL) solution to (75) of stirring.Then, on silicon-dioxide, above-mentioned reaction mixture is concentrated.Use toluene-ethyl acetate as elutriant gained resistates to be carried out column chromatography at 1: 1 and separate, be given 75 (0.150g, 96%) of colourless hard stock liquid.

NMR data (400MHz, CDCl ₃): 1H, δ 1.46-1.53 (m, 9H, C (CH ₃) ₃), 2.70 (OH-is less important for d, 0.3H), 3.26-3.46 (H-6A and OH-are main for m, 1.7H), 3.75-4.04 (m, 3H, H-1A, H-1B and H-6B), 4.29,4.34 (2d, 1H, H-3 ^*Less important and main), 4.43,4.50 (H-2 is less important and main for 2brs, 1H), 4.74 (m, 1H, H-4 ^*), 4.89,5.02 (2brs, 1H, H-5).

Step f)

Under 0 ℃, (0.099g adds trifluoroacetic acid (2mL) in methylene dichloride 0.40mmol) (2mL) solution, at room temperature stirs 35min then, concentrates then and (concentrates in 3 * 5mL) at toluene to (75).To gained resistates, I-hydroxybenzotriazole hydrate (0.067g, 0.44mmol), N-ethyl-N '-(3-dimethylaminopropyl) carbodiimide xHCl (0.084g, 0.44mmol) and N-(tertbutyloxycarbonyl)-L-leucine monohydrate (0.105g, 0.42mmol) DMF (4mL) suspension in add triethylamine (0.17mL, 1.2mmol), at room temperature its stirring is spent the night then.Then above-mentioned reaction is concentrated into half volume, with ethyl acetate (25mL) dilution, order with 10% aqueous citric acid solution (3 * 15mL) and the 1M sodium bicarbonate aqueous solution (3 * 15mL) washings are carried out drying (sodium sulfate), are filtered and concentrated.Use 3: 2 pairs of gained resistatess of ethyl acetate-toluene to carry out column chromatography and separate, be provided as (76) (0.137g, 95%) of colourless hard stock liquid.

NMR data (400MHz, CDCl ₃, the signal of selection): 1H, δ 0.89-1.01 (m, 6H, C (CH) ₂), 4.98,5.07 (H-5 is main and H-5 is less important for 2dd, 1H).

LR-MS:C ₁₇H ₃₀FN ₂O ₅Calculated value: 361.2. measured value: 361.1[M+H].

LR-MS:C ₂₁H ₂₆FN ₂O ₅: calculated value 405.2. measured value: 405.1[M+H].

The capping group coupling of P2-P1 structural unit and suitable haloalkylization and shown in above model compound, the P1 hydroxyl oxygen is changed into ketone.

Embodiment 5

Novel P2 structural unit

^*(+)-1,2-pair (2S, 5S)-diethylphosphonolanbenzene (cyclooctadiene) rhodium (I) fluoroform sulphonate

By J.Am.Chem.Soc., the method described in the Vol.103No.21981436-442 is prepared 1-methyl-cyclobutane carboxylate 1 by the cyclobutane carboxylate.

Under 0 ℃, under nitrogen atmosphere, 1-methyl-cyclobutane carboxylate 1 (1eq) is stirred in anhydrous THF.Portioning adds lithium aluminum hydride (1.5eq) in this solution, and at room temperature gained suspension is stirred 3 hours.The gained reaction mixture cools off on ice, handles and stirs 20 minutes down at 0 ℃ with 1MHCl (aqueous solution).Make gained solution by the diatomite pad, and gained filtrate is extracted in the ether.Gained organic phase MgSO ₄Dry, filter and concentrate in a vacuum, thereby provide (1-methyl-cyclobutyl)-methyl alcohol 2.

The diatomite of pyridinium chlorochromate (1.25eq) and identical weight is absorbed in the anhydrous methylene chloride as suspension.In this suspension, drip the anhydrous methylene chloride solution that adds compound 2 (1eq), and at room temperature the multiphase mixture that obtains was stirred 3 hours.Make reaction mixture pass through the silicon-dioxide pad, with 19: 1 isohexanes: eluent ethyl acetate, thus 1-methyl cyclobutane formaldehyde 3 provided.

Under agitation compound 3 (1eq) is dissolved in the anhydrous methylene chloride, and in this solution, adds Boc-phosphorus glycine trimethyl (0.5eq) and 1,8-diazabicylo [5.4.0]-7-alkene (1.2eq).Under nitrogen, gained solution stirred at ambient temperature spend the night.Reaction mixture is distributed in methylene dichloride and 1M HCl (aqueous solution) in succession, saturated NaHCO ₃Among (aqueous solution) and the saturated NaCl (aqueous solution).Organic layer MgSO ₄Dry, filter and under vacuum, concentrate.By flash column chromatography gained oil is carried out purifying,, thereby provide 2-t-butoxycarbonyl amino-3-(1-methyl-cyclobutyl)-methyl acrylate 4 with the dichloromethane solution wash-out of 1% methyl alcohol.

Compound 4 is dissolved in the anhydrous methanol and with nitrogen outgases.With (+)-1, and 2-two (2S, 5S)-diethylphosphonolanbenzene (cyclooctadiene) rhodium (I) fluoroform sulphonate adds wherein and continuation outgased 10 minutes.(4bar) should react jolting 48 hours under hydrogen atmosphere.In a vacuum solution is concentrated and carry out purifying, use the methylene dichloride wash-out, thereby provide 2S-t-butoxycarbonyl amino-3-(1-methyl-cyclobutyl)-methyl propionate 5 by flash chromatography.

HPLC retention time 5.88min (215 and 254nm under monitor)

HPLC uses Synergy Max RP 80 μ m 50 * 4.6mm post, 10 → 90%6min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A), flow velocity 2ml/min.

MS[M+H] ⁺272.08(20％)[M-Boc+H] ⁺172.06(100％)

Electrospray ionization is with acetonitrile/ammonium formiate buffer solution elution.

¹H NMR(400MHz，CDCl ₃.4.89-4.79(1H，m)4.33-4.27(1H，m)3.71(3H，s)1.98-1.62(8H，m)1.42(9H，s)1.22(3H，s)

Embodiment 6

(S)-2-[(S)-and 1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-valeric acid.

This title compound such as Li, people BiooRg.Med.Chem.Lett.200616 such as C.S. are prepared shown in 1985.

Embodiment 7

(S)-4-isobutyl--2-trifluoromethyl-oxazolidines

This title compound such as Ishii are prepared shown in the people Synlett 1997,1381 such as A..

Embodiment 8

[(S)-1-(tertiary butyl-dimethyl-silane oxygen ylmethyl)-3-methyl-butyl]-[2,2,2-three fluoro-second-(E)-subunit]-amine

This title compound such as Li, people BiooRg.Med.Chem.Lett.200616 such as C.S. are prepared shown in 1985.

Embodiment 9

(3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-; Muriate

This title compound goes protection by the structural unit to embodiment 1 and is prepared with the salt acid treatment.

Embodiment 10

(S)-and 2-[1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-penta-1-alcohol

Under-78 ℃, at N ₂In, to 1 of stirring, the 4-dibromobenzene (1.79g, and the hexane solution of dropping adding 1.6M butyllithium in anhydrous THF (5ml) solution 7.61mmol) (4.5ml, 7.61mmol).Under-78 ℃, gained solution was further stirred 20 minutes, after this, (0.5g, anhydrous THF (5ml) solution 2.54mmol) joins in the lithium aryl solution with (S)-4-isobutyl--2-trifluoromethyl-oxazolidine.Under-78 ℃, continue to stir 1 hour.Reaction mixture is with 5ml 2M hydrochloric acid soln quencher and make mixture be warming up to room temperature.Gained solution alkalize with 10ml sodium hydroxide and gained solution (2 * 30ml) extract, and the organic constituent that is combined carries out drying (MgSO with ethyl acetate ₄) and concentrate in a vacuum.(ethyl acetate: isohexane 1: 4) products obtained therefrom is carried out purifying, thereby obtain the title product (0.620g, 53%) into yellow oil, it is 2: 1 mixtures of diastereomer by flash column chromatography.MS M+H 355, retention time 4.9﹠amp; 5.1min the anti-phase 50mm*4.6mm i.d. of 10-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 11

(S)-and 4-methyl-2-[2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-alcohol

To (the S)-2-[1-(4-bromo-phenyl)-2 that stirs, 2,2-three fluoro-ethylaminos]-4-methyl-penta-1-alcohol (0.1g, 0.28mmol) DMF (5ml) solution in add (4-methylsulfonyl phenyl) boric acid (0.067g, 0.34mmol), sodium carbonate solution (0.090g, 0.85mmol, in 5ml water) and [1,1 '-two (diphenyl phosphine) ferrocene] palladium (II) muriate and CH ₂Cl ₂1: 1 title complex (0.023g, 0.03mmol).Under 80 ℃, above-mentioned gained solution was heated 60 minutes.Make above-mentioned reaction mixture be cooled to room temperature and use CH ₂Cl ₂(20ml) dilute.With separating organic matters, carry out drying (MgSO ₄) and concentrate in a vacuum.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain the title product (0.087g, 53%) into yellow solid, it is 2: 1 mixtures of diastereomer.Retention time 4.2﹠amp; The anti-phase 50mm*4.6mm i.d. of 4.3min10-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 12

(S)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-valeric acid;

(S)-the 4-methyl-2-[(R)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-valeric acid

To the periodic acid that stirs (5.7g, add in acetonitrile 4.19mmol) (55ml) suspension chromic oxide (VI) (11mg, 0.011mmol) and water (0.25ml).At room temperature with gained suspension stirred for several hour; on ice bath, it is cooled to 0 ℃ then; and with (S)-4-methyl-2-[2; 2; 2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-alcohol (1.8g; 4.19mmol) join in the 10ml acetonitrile, continue down to stir 30 minutes at 0 ℃.Reaction is poured into acidifying Na ₂HPO ₄In the solution (12.0g in 200ml water, regulates pH value 3 with dense HCl), use then ether (3 * 100ml) extract, the organic extraction of merging use continuously salt solution (1 * 100ml), sodium sulfite solution (1 * 100ml) and salt solution (1 * 100ml) washs.Organic extraction is carried out drying (Na ₂SO ₄) and concentrate, thereby being given the title product (1.3g, 53%) of yellow oil, it is 2: 1 mixtures of diastereomer.Retention time 4.2﹠amp; 4.4min the anti-phase 50mm*4.6mm i.d. of 10-90MeCN:0.05%TFA 6min gradient C12 post.The diastereo-isomerism mixture of product is further by preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post; 30 → 90%15min solution B gradient (acetonitrile solution of solution A=0.1%TFA aqueous solution and solution B=10%A) flow velocity 18ml/min] carry out purifying; thereby the ratio of obtaining is two kinds of isolating diastereomers for the title compound of white solid of 2: 1; (S)-the 4-methyl-2-[(S)-2; 2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-valeric acid

(0.192g) M+H 444, (S)-and the 4-methyl-2-[(R)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-valeric acid (0.120g) M+H 444.

Embodiment 13

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(R)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone

To stir (3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-; Muriate (0.05g, CH 0.30mmol) ₂Cl ₂(2ml) and (S)-the 4-methyl-2-[(R)-2; 2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-(0.120g 0.27mmol) adds dicyclohexylcarbodiimide (0.12g to valeric acid in the solution; 0.54mmol) and diisopropylethylamine (0.047ml, 0.27mmol).At room temperature with above-mentioned gained solution stirring 60 minutes.To react then filtration over celite and then the gained organism with 2M HCl (1 * 2ml), saturated NaHCO ₃(drying (MgSO is carried out in 1 * 2ml) washing ₄) and concentrate in a vacuum.By flash column chromatography (isohexane: ethyl acetate, the 25-100% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.061g into white foam, 39%), M+H 573, the anti-phase 50mm*4.6mm i.d. of retention time 5.8min 10-90MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 13A

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone

To stir (3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-; Muriate (0.08g, CH 0.47mmol) ₂Cl ₂(2ml) and (S)-the 4-methyl-2-[(S)-2; 2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-(0.19g 0.43mmol) adds dicyclohexylcarbodiimide (0.18g to valeric acid in the solution; 0.86mmol) and diisopropylethylamine (0.074ml, 0.43mmol).At room temperature with above-mentioned gained solution stirring 60 minutes.To react then filtration over celite and then the gained organism with 2M HCl (1 * 2ml), saturated NaHCO ₃(drying (MgSO is carried out in 1 * 2ml) washing ₄) and concentrate in a vacuum.By flash column chromatography (isohexane: ethyl acetate, the 25-100% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.072g into white foam, 29%), M+H 573, the anti-phase 50mm*4.6mm i.d. of retention time 5.8min 10-90MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 14

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

To (the S)-1-((3R, 3aR, the 6S that stir; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2; 2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.072g, CH 0.13mmol) ₂Cl ₂(5ml) add in the solution Dess Martin cross iodine alkane (0.11g, 0.25mmol).At room temperature with above-mentioned gained solution stirring 2 hours.Gained reaction mixture CH ₂Cl ₂(20ml) saturated NaHCO is used in dilution ₃(drying (MgSO is carried out in 2 * 10ml) washings ₄) and concentrate in a vacuum.By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain title compound (0.038g into white solid, 52%), is and its mixture of ketone hydrate.MS M+H 571，M+H ₂O+H 589。Retention time 5.5﹠amp; 5.9min the anti-phase 50mm*4.6mmi.d. post of 10-90MeCN:0.05%TFA 6min gradient C12

Embodiment 15

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(R)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R; 3aR; 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3; 2-b] pyrroles-4-yl)-the 4-methyl-2-[(R)-2; 2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.061g, 0.11mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtaining title compound (0.021g, 34%), is the mixture of ketone hydrate with it.MS M+H 571, M+H ₂O+H 589. retention time 5.3﹠amp; 5.8min the anti-phase 50mm*4.6mmi.d. post of 10-90MeCN:0.05%TFA 6min gradient C12

Embodiment 16

(S)-2-[(S)-and 1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-4-methyl-penta-1-ketone

To stir (3S, 3aS, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-; Muriate (2.250g, 12.25mmol) DMF (40ml) and (S)-2-[(S)-1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-valeric acid (4.1g, 11.14mmol) add HATU (5.08g in the solution, 13.36mmol) and diisopropylethylamine (5.8mls, 33.41mmol).At room temperature above-mentioned gained solution stirring is spent the night, in a vacuum it is concentrated then.The gained resistates is dispersed in the water (50ml) and (2 * 150ml) extract with ethyl acetate.The organic constituent that merges is with saturated sodium bicarbonate solution (1 * 100ml) washing and carry out drying and concentrate with sal epsom.By flash column chromatography (isohexane: ethyl acetate, the 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (2.95g into yellow oil, 53%) MS M+H 497, the anti-phase 50mm*4.6mm i.d. of retention time 5.8min 10-90MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 17

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone

To (S)-the 2-[(S)-1-(4-bromo-phenyl)-2 that stirs; 2; 2-three fluoro-ethylaminos]-1-((3R; 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3; 2-b] pyrroles-4-yl)-4-methyl-penta-1-ketone (0.1g; 0.20mmol) DMF (1ml) solution in add (3-methylsulfonyl phenyl) boric acid (0.044g, 0.22mmol), 2M sodium carbonate solution (1ml) and [1,1 '-two (diphenyl phosphine) ferrocene] palladium (II) muriate and CH ₂Cl ₂(0.016g, 1: 1 title complex 0.02mmol).Gained solution is sealed in the test tube, and in microwave, is heated to 160 ℃, kept 5 minutes.Make above-mentioned reaction mixture be cooled to room temperature and use CH ₂Cl ₂: H ₂O (1: 1,10ml) dilute.With separating organic matters, carry out drying (MgSO ₄) and concentrate in a vacuum.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.048g, 42%) into yellow oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 573. retention time 5.4min 10-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 18

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Embodiment 14 described technologies are used for (S)-1-((3R; 3aR; 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3; 2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2; 2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.048g, 0.11mmol).By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby provides title product (0.013g, 29%), for its mixture of ketone hydrate.MS M+H 571, M+H ₂O+H 589. retention time 4.2﹠amp; 4.7min the anti-phase 50mm*4.6mm i.d. of 10-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 19

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-fluoro-biphenyl-4-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 17 is used for (S)-2-[(S)-1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 4-fluorophenyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.069g, 45%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 513. retention time 5.6min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 20

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-fluoro-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Embodiment 14 described technologies are used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-fluoro-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.069g, 0.13mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.036g, 34%), is and its mixture of ketone hydrate.MS M+H 511, M+H ₂O+H 529. retention time 5.5﹠amp; 6.0min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 21

4 '-(S)-2,2,2-three fluoro-1-[(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-carbonyl)-3-methyl-butyl amino]-ethyl }-biphenyl-4-nitrile

Embodiment 17 described technologies are used for (S)-2-[(S)-1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 4-cyano-phenyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.054g, 35%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 513. retention time 5.2min 30-90MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 22

4 '-(S)-2,2,2-three fluoro-1-[(S)-1-((3aS, 6S, 6aS)-6-fluoro-3-oxo-six hydrogen-furo [3,2-b] pyrroles-4-carbonyl)-3-methyl-butyl amino]-ethyl }-biphenyl-4-nitrile

Technology described in the embodiment 14 is used for 4 '-(S)-2,2,2-three fluoro-1-[(S)-1-((3aS, 6S, 6aS)-6-fluoro-3-oxo-six hydrogen-furo [3,2-b] pyrroles-4-carbonyl)-3-methyl-butyl amino]-ethyl }-biphenyl-4-nitrile (0.054g, 0.13mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.026g, 34%), is and its mixture of ketone hydrate.MS M+H 518, M+H ₂O+H536. retention time 5.1﹠amp; 5.6min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 23

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-trifluoromethyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 17 is used for (S)-2-[(S)-1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 4-trifluorophenyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.086g, 51%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 563. retention time 5.9min30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 24

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-trifluoromethyl-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-trifluoromethyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.086g, 0.15mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.056g, 34%), is and its mixture of ketone hydrate.MS M+H 561, M+H ₂O+H579. retention time 6.0﹠amp; 6.5min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 25

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(2 '-fluoro-biphenyl-4-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 17 is used at dme: ethanol (1: 1, (S)-2-[(S)-1-1ml) (4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.13g is 0.26mmol) with 2-fluorophenyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.05g, 38%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 513. retention time 5.4min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 26

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(2 '-fluoro-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(2 '-fluoro-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.05g, 0.10mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.036g, 72%), is and its mixture of ketone hydrate.MS M+H 511, M+H ₂O+H 529. retention time 5.5﹠amp; 6.0min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 27

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl sulfane base-biphenyl-4-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 17 is used at dme: ethanol (1: 1, (S)-2-[(S)-1-1ml) (4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.13g is 0.26mmol) with 4-methylthio phenyl ylboronic acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.07g, 50%) into oil.The anti-phase 50mm*46mm i.d. of MS M+H541. retention time 5.8min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 28

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl sulfane base-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl sulfane base-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.07g, 0.13mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.007g, 10%), is and its mixture of ketone hydrate.MS M+H 539, M+H ₂O+H557. retention time 5.7﹠amp; 6.3min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 29

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl sulfoxide-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl sulfane base-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.07g, 0.13mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.027g, 39%), is and its mixture of ketone hydrate.MS M+H 555, M+H ₂O+H573. retention time 3.9﹠amp; 4.5min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 30

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methoxyl group-biphenyl-4-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 17 is used at dme: ethanol (1: 1, (S)-2-[(S)-1-1ml) (4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.13g is 0.26mmol) with 4-anisole ylboronic acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.049g, 36%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H525. retention time 5.6min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 31

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methoxyl group-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methoxyl group-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.049g, 0.09mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.033g, 68%), is and its mixture of ketone hydrate.MS M+H 523, M+H ₂O+H541. retention time 5.3﹠amp; 5.9min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 32

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 17 is used at dme: ethanol (1: 1, (S)-2-[(S)-1-1ml) (4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.13g is 0.26mmol) with 4-aminomethyl phenyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.051g, 38%) into oil.The anti-phase 50mm*46mm i.d. of MS M+H509. retention time 5.8min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 33

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.05g, 0.10mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.033g, 66%), is and its mixture of ketone hydrate.MS M+H 507, M+H ₂O+H 525. retention time 5.8﹠amp; 6.4min the anti-phase 50mm*4.6mmi.d. post of 30-90MeCN:0.05%TFA 6min gradient C12

Embodiment 34

[(S)-and 1-(3-bromo-phenyl)-2,2,2-three fluoro-ethyls]-[(S)-1-(tertiary butyl-dimethyl-silane oxygen ylmethyl)-3-methyl-butyl]-amine

Under-78 ℃, at N ₂In, to 1 of stirring, the 3-dibromobenzene (9.1g, and the hexane solution of dropping adding 1.6M butyllithium in anhydrous THF (100ml) solution 38.53mmol) (24.1ml, 38.53mmol).Under-78 ℃, gained solution was stirred 20 minutes in addition, to after this [(S)-and 1-(tertiary butyl-dimethyl-silane oxygen ylmethyl)-3-methyl-butyl]-[2,2,2-three fluoro-second-(E)-subunit]-(4.0g, anhydrous THF (10ml) solution 12.84mmol) joins in the solution of lithium aryl amine.Under-78 ℃, continue to stir 1 hour.Reaction mixture is with 50ml 2M hydrochloric acid soln quencher and make mixture be warming up to room temperature.Gained solution alkalize with 100ml sodium hydroxide and gained solution (2 * 100ml) extract, and the organic constituent that is combined carries out drying (MgSO with ethyl acetate ₄) and concentrate in a vacuum.By anti-phase C18 column chromatography (H ₂O: MeCN, the 50-100% gradient) products obtained therefrom is carried out purifying, thus obtain title product (2.55g 42%) MS M+H 468, the anti-phase 50mm*4.6mm i.d. of retention time 8.3min 50-97MeCN:0.05%TFA 6min gradient C12 post into yellow oil

Embodiment 35

(S)-2-[(S)-and 1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-penta-1-alcohol

To stir [(S)-1-(3-bromo-phenyl)-2,2,2-three fluoro-ethyls]-[(S)-1-(tertiary butyl-dimethyl-silane oxygen ylmethyl)-3-methyl-butyl]-(2.55g adds concentrated hydrochloric acid (1ml) in methyl alcohol 5.4mmol) (60ml) solution to amine, and heats 18 hours down at 0 ℃.Make reaction cooling and in a vacuum it is concentrated.By flash column chromatography (isohexane: ethyl acetate, 1-33% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (1.57g, 82%) into yellow oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 354. retention time 4.1min 30-97MeCN:0.05%TFA 6min gradient C12 post

Embodiment 36

(S)-2-[(S)-and 1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-valeric acid

To the periodic acid that stirs (11.5g, add in acetonitrile 50.82mmol) (100ml) suspension chromic oxide (VI) (23mg, 0.023mmol) and water (0.25ml).At room temperature with gained suspension stirred for several hour, on ice bath, it is cooled to 0 ℃ then, with (S)-2-[(S)-1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-penta-1-alcohol (1.5g, 4.19mmol) join in the 10ml acetonitrile, continue down to stir 30 minutes at 0 ℃.Reaction is poured into acidifying Na ₂HPO ₄In the solution (12.0g in 200ml water, regulates pH value 3 with dense HCl), use then ether (3 * 100ml) extract, the organic extraction of merging use in proper order salt solution (1 * 100ml), sodium sulfite solution (1 * 100ml) and salt solution (1 * 100ml) washs.The gained organic extraction is carried out drying (Na ₂SO ₄) and concentrate, thereby be given title product (1.3g, 85%) the MS M+H 368 of yellow oil, the anti-phase 50mm*4.6mm i.d. of retention time 5.0min 30-97MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 37

(S)-2-[(S)-and 1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-4-methyl-penta-1-ketone

To stir (3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-; Muriate (0.71g, DMF 3.88mmol) (10ml) and (S)-2-[(S)-1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-valeric acid (1.3g, 3.53mmol) add in the solution HATU (1.6g, 4.24mmol) and diisopropylethylamine (1.8ml, 10.59mmol).At room temperature above-mentioned gained solution stirring is spent the night, in a vacuum it is concentrated then.Then the gained resistates is dispersed in the water (50ml) and (2 * 150ml) extract with ethyl acetate.The organic constituent that merges is with saturated sodium bicarbonate solution (1 * 100ml) washing and carry out drying and concentrate with sal epsom.By anti-phase C18 column chromatography (H ₂O: acetonitrile, 30-90% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.96g, 55%) MS M+H 497, the anti-phase 50mm*4.6mm i.d. of retention time 4.7min 30-90MeCN:0.05%TFA 6min gradient C12 post into yellow oil.

Embodiment 38

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl-biphenyl-3-yl)-ethylamino]-penta-1-ketone

To (S)-the 2-[(S)-1-(3-bromo-phenyl)-2 that stirs; 2; 2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3; 2-b] pyrroles-4-yl)-4-methyl-penta-1-ketone (0.15g, dimethyl ether 0.30mmol): ethanol (1: 1,1ml) solution adds 4-methylsulfonyl phenyl-boron dihydroxide (0.044g; 0.22mmol), the tetrakis triphenylphosphine palladium (0) of 2M sodium carbonate solution (1ml) and polymkeric substance load (0.15g, 0.015mmol).Gained solution is sealed in the test tube, and in microwave, is heated to 160 ℃, kept 5 minutes.Make above-mentioned reaction mixture be cooled to room temperature, use CH ₂Cl ₂: H ₂O (1: 1,10ml) dilute and filter.With separating organic matters, carry out drying (MgSO ₄) and concentrate in a vacuum.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.071g, 48%) into clean oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 509. retention time 5.5min 10-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 39

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl-biphenyl-3-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methyl-biphenyl-3-yl)-ethylamino]-penta-1-ketone (0.072g, 0.14mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.033g, 46%), is and its mixture of ketone hydrate.MS M+H 507, M+H ₂O+H525. retention time 5.9﹠amp; 6.6min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 40

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-3-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 38 is used for (S)-2-[(S)-1-(3-bromo-phenyl)-2; 2; 2-three fluoro-ethylaminos]-1-((3R; 3aR; 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 4-methylsulfonyl phenyl-boron dihydroxide for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-100% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.07g, 41%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 573. retention time 4.3min30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 41

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-3-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R; 3aR; 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3; 2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2; 2,2-three fluoro-1-(4 '-methylsulfonyl-biphenyl-3-yl)-ethylamino]-penta-1-ketone (0.07g, 0.12mmol).By preparation-HPLC[Phenomenex Synergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.027g, 39%), is and its mixture of ketone hydrate.MS M+H 571, M+H ₂O+H589. retention time 5.2﹠amp; 5.4min the anti-phase 50mm*4.6mm i.d. of 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 42

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-3-yl)-ethylamino]-penta-1-ketone

Embodiment 38 described technologies are used for (S)-2-[(S)-1-(3-bromo-phenyl)-2; 2; 2-three fluoro-ethylaminos]-1-((3R; 3aR; 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 3-methylsulfonyl phenyl-boron dihydroxide for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-100% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.1g, 60%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 573. retention time 4.4min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 43

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-3-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R; 3aR; 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3; 2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2; 2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-3-yl)-ethylamino]-penta-1-ketone (0.1g, 0.18mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.051g, 51%), is and its mixture of ketone hydrate.MS M+H 571, M+H ₂O+H 589. retention time 5.3﹠amp; 5.5min 30-90MeCN:0.05%TFA, the anti-phase 50mm*4.6mm i.d. of 6min gradient C12 post

Embodiment 44

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(2 '-fluoro-biphenyl-3-yl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 38 is used for (S)-2-[(S)-1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 2-fluorophenyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.11g, 69%) into oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 513. retention time 5.1min 30-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 45

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(2 '-fluoro-biphenyl-3-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(2 '-fluoro-biphenyl-3-yl)-ethylamino]-penta-1-ketone (0.11g, 0.21mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.054g, 51%), is and its mixture of ketone hydrate.MS M+H 511, M+H ₂O+H 529. retention time 5.4﹠amp; 6.1min 30-90MeCN:0.05%TFA, the anti-phase 50mm*4.6mmi.d. post of 6min gradient C12

Embodiment 46

(S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(3-pyridin-4-yl-phenyl)-ethylamino]-penta-1-ketone

Technology described in the embodiment 38 is used for (S)-2-[(S)-1-(3-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-(0.15g is 0.30mmol) with 4-pyridyl boric acid for 4-methyl-penta-1-ketone.By flash column chromatography (isohexane: ethyl acetate, 5-100% gradient) products obtained therefrom is carried out purifying, thereby obtain title product (0.071g, 47%) into oil.MS M+H 496. retention time 3.7min 30-90MeCN:10mM (NH ₃) ₂CO ₃The anti-phase 50mm*4.6mm i.d. of 6min gradient C12 post

Embodiment 47

(3aS, 6S, 6aS)-6-fluoro-4-{ (S)-4-methyl-2-[(S)-2,2,2-three fluoro-1-(3-pyridin-4-yl-phenyl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-the 4-methyl-2-[(S)-2,2,2-three fluoro-1-(3-pyridin-4-yl-phenyl)-ethylamino]-penta-1-ketone (0.071g, 0.14mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.008g, 11%), is and its mixture of ketone hydrate.MS M+H 494, M+H ₂O+H 512. retention time 4.0﹠amp; 4.6min 30-90MeCN:10mM (NH ₃) ₂CO ₃, the anti-phase 50mm*4.6mm i.d. of 6min gradient C12 post

Embodiment 48

1-(4-bromo-thiazolyl-2-yl)-4-methyl-piperazine

People such as title compound such as Palmer J.Med.Chem.2005,48, be prepared shown in the 7520-7534.

Embodiment 49

(3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-benzyl carboxylate

This title compound carries out conventional deprotection/protection by the structural unit by embodiment 1 and is prepared.

Embodiment 50

(S)-2-[(S)-and 1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-isopropyl isovalerate

To (S)-the 2-[(S)-1-(4-bromo-phenyl)-2,2 that stirs, 2-three fluoro-ethylaminos]-(1.80g, Virahol 4.9mmol) (100ml) solution joins in the vitriol oil (2ml) 4-methyl-valeric acid.Under 80 ℃, above-mentioned reaction mixture was heated 4 hours.Make the reaction mixture cooling, concentrate in a vacuum and the gained oil content is dispersed in CH ₂Cl ₂(100ml), use saturated NaHCO ₃(2 * 50ml) wash, carry out drying (MgSO ₄) and concentrate in a vacuum, thereby obtain title compound (1.77g, 88%) into brown oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 412. retention time 6.6min30-97MeCN:0.05%TFA 6min gradient C12 post

Embodiment 51

(S)-and 4-methyl-2-{ (S)-2,2,2-three fluoro-1-[4-(4,4,5,5-tetramethyl--[1,3,2] dioxy boron penta ring-2-yl)-phenyl]-ethylamino }-isopropyl isovalerate

To (S)-the 2-[(S)-1-(4-bromo-phenyl)-2 that stirs, 2,2-three fluoro-ethylaminos]-(2.2g adds two (pinacolyl) borine (2.0g to 4-methyl-isopropyl isovalerate in DMF 5.36mmol) (30ml) solution, 8.04mmol), potassium acetate (1.6g, 16.1mmol) and and CH ₂Cl ₂1: 1 title complex of [1,1 '-two (diphenyl phosphine) ferrocene] palladium (II) muriate (0.438g, 0.54mmol).Gained solution is sealed in the test tube, and in microwave, is heated to 160 ℃, kept 20 minutes.Make reaction mixture be cooled to room temperature, and filter short silica column with ethyl acetate (500ml) washing.In a vacuum gained solution is concentrated and by anti-phase C18 column chromatography (H ₂O: MeCN, the 50-100% gradient) the thick product of gained is carried out purifying, thus obtain title compound (0.920g, 38%) into brown oil.The anti-phase 50mm*4.6mm i.d. of MS M+H 458. retention time 4.6min 70-97MeCN:0.05%TFA 6min gradient C12 post

Embodiment 52

(S)-4-methyl-2-((S)-2,2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazolyl-4-yl]-phenyl }-ethylamino)-isopropyl isovalerate

To (the S)-4-methyl-2-{ (S)-2,2 that stirs, 2-three fluoro-1-[4-(4,4,5,5-tetramethyl--[1,3,2] dioxy boron penta ring-2-yl)-phenyl]-ethylamino }-isopropyl isovalerate (0.72g, DMF 1.57mmol): H ₂O (1: 1,20ml) add in the solution 1-(4-bromo-thiazol-2-yl)-4-methyl-piperazine (0.5g, 1.89mmol), yellow soda ash (0.2g, 1.89mmol) and and CH ₂Cl ₂1: 1 title complex of [1,1 '-two (diphenyl phosphine) ferrocene] palladium (II) muriate (0.129g, 0.16mmol).Gained solution is sealed in the test tube, and in microwave, is heated to 160 ℃, kept 20 minutes.Make above-mentioned reaction mixture cooling and use CH ₂Cl ₂(100ml) dilute.Drying (MgSO is separated, carried out to organic phase ₄) and concentrate in a vacuum.By flash column chromatography (ethyl acetate: MeOH, 9: 1) the thick product of gained is carried out purifying, thereby obtain being garnet solid title product (0.150g, 13%).MS M+H 513. retention time 4.0min 50-9710mM (NH ₃) ₂CO ₃: the anti-phase 50mm*4.6mm i.d. of MeCN 6min gradient C12 post

Embodiment 53

(S)-4-methyl-2-((S)-2,2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazole-4-yl]-phenyl }-ethylamino)-valeric acid; Hydrochloride

To the 2M hydrochloric acid that stirs with diox (1: 1,10ml) add (S)-4-methyl-2-((S)-2 in the mixture, 2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazolyl-4-yl]-phenyl }-ethylamino)-isopropyl isovalerate (0.15g, 0.29mmol).Then, with gained solution heating 20 hours, be concentrated into drying then in a vacuum, thereby be given dun solid title compound (0.14g, 98%) under 100 ℃, it does not need to be further purified and can use.MS M-H 469.

Embodiment 54

(3aS, 6S, 6aS)-6-fluoro-3-oxo-six hydrogen-furo [3,2-b] pyrroles-4-benzyl carboxylate

To stir (3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-benzyl carboxylate (1.25g, CH 4.44mmol) ₂Cl ₂(50ml) add in the solution Dess-Martin cross iodine alkane (2.1g, 4.44mmol).At room temperature with above-mentioned gained solution stirring 2 hours.The saturated NaHCO of above-mentioned reaction mixture ₃And 10%Na ₂S ₂O ₃(1: 1,100ml) quencher was carried out drying (MgSO to organic phase to the mixture of solution ₄) and concentrate in a vacuum.By flash column chromatography (isohexane: ethyl acetate 1: 1) the thick product of gained is carried out purifying, thereby be given the title compound (1.15g, 92%) of clean oil.MS M+H 280, TLC R _f0.2 (isohexane: ethyl acetate 1: 2)

Embodiment 55

(3aS, 6S, 6aS)-and 6-fluoro-3,3-dimethoxy-six hydrogen-furo [3,2-b] pyrroles-4-benzyl carboxylate

To stir (3aS, 6S, 6aS)-(1.15g adds trimethyl orthoformate (5ml) and tosic acid (0.02g) to 6-fluoro-3-oxo-six hydrogen-furo [3,2-b] pyrroles-4-benzyl carboxylate in methyl alcohol 4.1mmol) (10ml) solution.Under 60 ℃,, make its cooling then and concentrate in a vacuum gained solution stirring 3 hours.By flash column chromatography (isohexane: ethyl acetate, 5-66% gradient) the thick product of gained is carried out purifying, thereby be given the title product (1.07g, 80%) of clean oil.MS M+H 326, TLC R _f0.2 (isohexane: ethyl acetate 1: 4)

Embodiment 56

(3aS, 6S, 6aS)-and 6-fluoro-3,3-dimethoxy-six hydrogen-furo [3,2-b] pyrroles

On the H-Cube hydrogenator, make (3aS, 6S, 6aS)-6-fluoro-3,3-dimethoxy-six hydrogen-furo [3,2-b] pyrroles-4-benzyl carboxylate (0.07g, methyl alcohol 0.22mmol) (10ml) solution with the flow velocity of 1ml/min by containing the cylinder of 10%Pd/C (10mg).Gained solution is concentrated, thereby be given the title product (0.042g, 99%) of clean oil.MS M+H 192.

Embodiment 57

(S)-1-((3aS, 6S, 6aS)-and 6-fluoro-3,3-dimethoxy-six hydrogen-furo [3,2-b] pyrroles-4-yl)-4-methyl-2-((S)-2,2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazole-4-yl]-phenyl }-ethylamino)-penta-1-ketone

To (the 3aS that stirs, 6S, 6aS)-6-fluoro-3,3-dimethoxy-six hydrogen-furo [3,2-b] pyrroles (0.084g, 0.44mmol) DMF (10ml) and (S)-4-methyl-2-((S)-2,2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazole-4-yl]-phenyl-ethylamino)-valeric acid; Hydrochloride (0.14g, add in solution 0.29mmol) HATU (0.17g, 0.44mmol) and diisopropylethylamine (0.15ml, 0.88mmol).At room temperature above-mentioned gained solution stirring is spent the night, in a vacuum it is concentrated then.Then, the gained resistates is dispersed in CH ₂Cl ₂(20ml) neutralization with saturated sodium bicarbonate solution (1 * 10ml) washing, and with dried over mgso with concentrate.By anti-phase C18 column chromatography (H ₂O: acetonitrile, 30-90% gradient) products obtained therefrom is carried out purifying, thereby obtain title compound (0.39g, 55%) the MS M+H 644 into light brown oil, retention time 4.7min 30-9710mM (NH ₃) ₂CO ₃: the anti-phase 50mm*4.6mm i.d. of MeCN 6min gradient C12 post.

Embodiment 58

(3aS.6S, 6aS)-6-fluoro-4-[(S)-4-methyl-2-((S)-2,2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazole-4-yl]-phenyl }-ethylamino)-pentanoyl]-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

To (S)-1-((3aS that stirs, 6S, 6aS)-6-fluoro-3,3-dimethoxy-six hydrogen-furo [3,2-b] pyrroles-4-yl)-4-methyl-2-((S)-2,2,2-three fluoro-1-{4-[2-(4-methyl-piperazine-1-yl)-thiazole-4-yl]-phenyl }-ethylamino)-(0.04g adds entry (0.2ml) in trifluoroacetic acid 0.062mmol) (1.8ml) solution to penta-1-ketone.At room temperature with above-mentioned gained solution stirring 24 hours.Reaction mixture CH ₂Cl ₂(5ml) dilution and utilize N ₂Air-flow evaporates solvent mixture.Then, the thick product of gained is dissolved in CH again ₂Cl ₂Neutralization 2M Na ₂CO ₃Solution (1ml) neutralization separates organic layer and concentrates in a vacuum.By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (solution A=10mM (NH ₃) ₂CO ₃The aqueous solution and the acetonitrile solution of solution B=10%A)] product is carried out purifying, thereby obtain to be white solid title compound (0.01g, 27%), be the mixture of ketone hydrate with it.MS M+H 598, M+H ₂O+H 616. retention time 3.2﹠amp; 3.8min 30-9710mM (NH ₃) ₂CO ₃: the anti-phase 50mm*4.6mmi.d. post of MeCN 6min gradient C12

Embodiment 59

(S)-and 2-[1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-valeric acid

To the periodic acid that stirs (11.6g, add in acetonitrile 50.82mmol) (100ml) suspension chromic oxide (VI) (23mg, 0.023mmol) and water (0.5ml).At room temperature, on ice bath, it is cooled to 0 ℃ then, with (S)-2-[1-(4-bromo-phenyl)-2 with gained suspension stirred for several hour, 2,2-three fluoro-ethylaminos]-(1.5g 4.23mmol) joins in the 10ml acetonitrile 4-methyl-penta-1-alcohol, continues down to stir 30 minutes at 0 ℃.Reaction is poured into acidifying Na ₂HPO ₄In the solution (12.0g in 200ml water, regulates pH value 3 with dense HCl), use then ether (3 * 100ml) extract, the organic extraction of merging use continuously salt solution (1 * 100ml), sodium sulfite solution (1 * 100ml) and salt solution (1 * 100ml) washs.Organic extraction is carried out drying (Na ₂SO ₄) and concentrate, thereby being given the title compound (1.61g, 97%) of yellow oil, it is 2: 1 mixtures of diastereomer.The anti-phase 50mm*4.6mm i.d. of MS M+H 368. retention time 5.2min 10-90MeCN:0.05%TFA 6min gradient C12 post

Embodiment 60

(S)-and 2-[1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-1-((3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-4-methyl-penta-1-ketone

To stir (3R, 3aR, 6S, 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-; Muriate (0.09g, CH 0.60mmol) ₂Cl ₂(2ml) and (S)-2-[1-(4-bromo-phenyl)-2,2,2-three fluoro-ethylaminos]-4-methyl-valeric acid (0.2g, add in solution 0.54mmol) DCC (0.22g, 1.09mmol) and diisopropylethylamine (0.1ml, 0.54mmol).At room temperature above-mentioned gained solution stirring is spent the night, then with its filtration over celite and in a vacuum it is concentrated.By flash column chromatography (isohexane: ethyl acetate, 1: 1) the gained compound is carried out purifying, thereby obtain title compound (0.155 into yellow solid, 58%) MS M+H 497, the anti-phase 50mm*4.6mm i.d. of retention time 5.8min 10-90MeCN:0.05%TFA 6min gradient C12 post.

Embodiment 61

(3aS, 6S, 6aS)-and 6-fluoro-4-{ (S)-4-methyl-2-[2,2,2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-pentanoyl }-tetrahydrochysene-furo [3,2-b] pyrroles-3-ketone

Technology described in the embodiment 14 is used for (S)-1-((3R, 3aR, 6S; 6aS)-6-fluoro-3-hydroxyl-six hydrogen-furo [3,2-b] pyrroles-4-yl)-4-methyl-2-[2,2; 2-three fluoro-1-(3 '-methylsulfonyl-biphenyl-4-yl)-ethylamino]-penta-1-ketone (0.120g, 0.27mmol).By preparation-HPLC[PhenomenexSynergi C ₁₂10 μ m, 10 * 150mm post, 30 → 90%15min solution B gradient (aqueous solution of solution A=0.1%TFA and the acetonitrile solution of solution B=10%A) flow velocity 18ml/min] product is carried out purifying, thereby obtain to be the title compound of white solid (0.036g, 34%), is and its mixture of ketone hydrate.MS M+H 495, M+H ₂O+H 513. retention time 5.9﹠amp; 6.4min 10-90MeCN:0.05%TFA, the anti-phase 50mm*4.6mm i.d. of 6min gradient C12 post

Biology embodiment

The mensuration of cathepsin K proteolysis catalytic activity

The suitable mensuration of cathepsin K uses human reconstitution cell enzyme to carry out, such as described in PDB.

ID BC016058 standard; MRNA; HUM; 1699BP.

DE human tissue Proteinase K (pycnodysostosis), (cDNA clones MGC:23107 to mRNA

RX MEDLINE；.RX PUBMED；12477932.

DR RZPD；IRALp962G1234.

DR SWISS-PROT；P43235；

The reconstitution cell cathepsin K can be expressed in multiple commercially available expression system, comprises that intestinal bacteria, pichia belong to and the baculovirus system.The enzyme of purifying is removed presequence by ordinary method and is activated.

Measure the standard test condition of kinetic constant and use fluorescence peptide matrix, be generally H-D-Ala-Leu-Lys-AMC, and or at the 100mM Mes/Tris that contains 1mM EDTA and 10mM 2 mercapto ethanol, determine among the pH 7.0, perhaps at the 100mM sodium phosphate, imM EDTA, 0.1%PEG4000pH 6.5 or contain 5mM EDTA and the 100mM sodium acetate of 20mM halfcystine, carry out among the pH 5.5, in various situations, the optional 1M DTT that contains as stablizer.The enzyme concn of using is 5nM.The raw material matrix solution is prepared as 10mM concentration in DMSO.Screening is carried out under the fixedly substrate concn of 60 μ M, and detailed dynamics research uses the coubling dilution of the matrix of 250 μ M to carry out.In mensuration, total DMSO concentration remains on below 3%.All mensuration are all carried out at ambient temperature.Product fluorescence (excite under 390nm, launch under 460nm) uses Labsystems Fluoroskan Ascent fluorescent plate reader to monitor.The production development curve produces along with the formation of AMC product after 15 fens clock times.

Cathepsin S K _iMeasure

Measure the boc-Val-Leu-Lys-AMC fluorescence matrix of using the human tissue proteolytic enzyme S that expresses baculovirus and being obtained from Bachem and carry out with 384 orifice plate specifications, wherein 7 kinds of test compounds can parallelly with the positive control that contains known cathepsin S inhibitor contrast be carried out.

The matrix dilution

With 280 μ l/ holes with 12.5%DMSO join 96 deep hole polypropylene boards two row B-H capable in.The matrix in 70 μ l/ holes is joined among the capable A.The mensuration damping fluid (pH 6.5 for 100mM sodium phosphate, 100mM NaCl) in 2 * 250 μ l/ holes is joined among the capable A, mix, and along the extremely capable H of plate doubling dilution.

The inhibitor dilution.

The mensuration damping fluid in 100 μ l/ holes is joined among the row 2-5 and 7-12 in 4 row of polypropylene board at the bottom of the 96 hole V.The mensuration damping fluid in 200 μ l/ holes is joined in row 1 and 6.

First kind of test compounds that will prepare in DMSO joins in the row 1 of top line, generally so that 10～30 times of initial thick K that determine to be provided _iVolume provide.Thick K _iCalculate according to preliminary operation, wherein the 1mM boc-VLK-AMC (being diluted in 1/10 dilution of the DMSO solution of measuring the 10mM raw material in the damping fluid) with 10 μ l/ holes is assigned to 96 hole Microfluor ^TMThe capable B-H neutralization of plate is assigned among the capable A with 20 μ l/ holes.The various 10mM test compounds of 2 μ l are joined capable A, in each independent hole of row 1-10.The mensuration damping fluid that 90 μ l are contained 1mM DTT and 2nM cathepsin S is added in each hole of capable B-H and with 180 μ l and joins among the capable A.Use hyperchannel transfer pipet mixing row A and double dilution are expert among the G.Mix row H and on spectrophotofluorometer, carry out reading.Reading is prism (Prism) data of match competitive inhibition equation, and S=100 μ M and K are set _M=100 μ M, thus K obtained _iEstimated value, maximum maximum 100 μ M.

Second test compounds is joined in the row 6 of first trip, the 3rd test compounds is joined in the row 1 of second row, or the like.In the row 6 of footline, add 1 μ l contrast.Mix row 1 and the double row 5 that are diluted to.Mix row 6 and the double row 10 that are diluted to.

The multistage transfer pipet of 8-passage that use is set to 5 * 10 μ l is distributed to the matrix in 10 μ l/ holes in the 384 hole assay plate.First column distribution that matrix is diluted plate is to go in all row of the assay plate that A begins.It is capable that alternative will correctly be skipped in the most advanced and sophisticated interval of hyperchannel transfer pipet.Secondary series is distributed to go in all row that B begins.

The multistage transfer pipet of 12-passage that use is set to 4 * 10 μ l is distributed to the inhibitor in 10 μ l/ holes in the 384 hole assay plate.First row that inhibitor is diluted plate is distributed in the alternate row of the assay plate that begins with A1.The most advanced and sophisticated alternative row of will correctly skipping at interval of hyperchannel transfer pipet.Similarly, second, third and fourth line are distributed to respectively in the alternately row and column that begins with A2, B1 and B2.

Mix 20ml and measure damping fluid and 20 μ l 1M DTT.Add the abundant cathepsin S that is enough to provide the 2nM final concn.

Use is such as the divider of Multidrop 384, with 30 μ l/ holes join the institute of assay plate porose in, and on such as the spectrophotofluorometer of Ascent, carry out reading.

The fluorescence reading of the enzymatic lysis degree of reflection fluorescence matrix (excite with emission wavelength and be respectively 390nm and 460nm, use the bandpass filters setting) though be inhibitor, is the fit line speed in each hole.

Use SigmaPlot 2000, the institute's foraminous match speed match that makes various inhibitor is to the competitive inhibition equation, thus definite V, Km and Ki value.

The Ki of cathepsin L

Method with following correction is used to measure the Ki of cathepsin L.

Described enzyme is commercially available human tissue proteolytic enzyme L (for example Calbiochem).Matrix is for being obtained from the H-D-Val-Leu-Lys-AMC of Bahcem.Measuring damping fluid is the 100mM sodium acetate, 1mM EDTA, pH5.5.In measuring damping fluid, DMSO raw material (being 10mM in 100%DMSO) is diluted to 10%.Before being about to use, in measuring damping fluid, enzyme being prepared as 5nM concentration and adding the 1mM dithiothreitol (DTT).The 2ul10mM inhibitor that will prepare in 100%DMSO distributes among the A that enters a profession.10ul 50uM matrix (1/200 dilution of=10mM raw material in DMSO dilutes in measuring damping fluid)

Suppress research

Utilize said determination, use the test compounds of varied concentration that potential inhibitor is screened.Reaction obtains in matrix buffered soln and the inhibitor starting by enzyme is joined.K _iValue is calculated according to equation 1

$v_0=\frac{\mathrm{VS}}{K_M(1+\frac{I}{K_i})+S}---\left(1\right)$

V wherein ₀Be speed of response, V is a top speed, and S is the Michaelis constant K _MThe concentration of matrix and the concentration that I is inhibitor.

The representational compound of the present invention is measured cathepsin K usefulness and selectivity in said determination.Should be pointed out that cathepsin L and S data for the micromole, and compound resists cathepsin K so effectively and makes the result exist with nmole.

	Cathepsin K IC 50	The IC of cathepsin L 50	Cathepsin S IC 50
				Embodiment 14	29nM	100uM	36uM
Embodiment 18	56nM	94uM	19uM
				Embodiment 20	180nM	30uM	37uM
Embodiment 26	44nM	25uM	22uM
				Embodiment 28	51nM	5.5uM	10uM
Embodiment 29	18nM	180uM	27uM
				Embodiment 31	70nM	29uM	19uM
Embodiment 33	190nM	20uM	20uM
				Embodiment 58	＜5nM	139uM	188uM
Embodiment 61	1nM	6uM	25uM

Obviously, The compounds of this invention is very effective to cathepsin K, and the selectivity that the L-Cysteine HCL Anhydrous cathepsin L that is closely related and S are had at least 100 times.In addition, these compounds generally have good water-permeable (the following test) and other DMPK performance.

Perviousness

This examples measure inhibitor is by the transhipment effect of human gastrointestinal passage cell.The passage number that the utilization of this mensuration is known is 40～60 Caco-2 cell.

The summit is the transhipment in the outside on earth

Usually, every kind of compound will be tested in 2～4 holes.The outside, the end and topped hole are 10 μ M with the normal concentration that contains 1.5mL and 0.4mL transport buffer (TB) and test substances respectively.In addition, all test solns and damping fluid will contain 1%DMSO.Before test, make the transport plates precoating contain the developing medium 30 minutes of 10% serum, thereby avoid the non-specific combination with plastic material.After 21～28 days, cell is prepared for permeability test in the substratum on the screen support device.

Transport plates no 1 comprises 3 row, 4 holes of every row.Row 1 expression washings, row 2 " 30 minutes " and row 3 " 60 minutes ".Transport plates no 2 comprises 3 row, 4 holes of every row, and an expression row 4 " 90 minutes ", row 5 " 120 minutes " and residue row are not specified.

Developing medium is removed and insert is changed over to the washing row (No.1) of the transport plates (plate no.1) in 2 plates of no insert from topped hole, it is expert at and has used 1.5mL transport buffer (pH 7.4 for HBSS, 25mM HEPES) to prepare in 1～5.In A → B screening, also contain 1% bovine serum albumin among the TB of end apertura lateralis.

0.5mL transport buffer (pH 6.5 for HBSS, 25mM MES) is joined in the insert, and under 37 ℃, in the polymix vibrator, cell monolayer is cushioned 30 minutes in the transhipment buffer system.After with the buffer system balance, the Transepithelial resistance value (TEER) in each hole is measured by EVOM cutting adhesion instrument.The TEER value is generally 400～1000 Ω/hole (passage number that depends on application).

(TB, pH 6.5) removes from the top side with transport buffer, and insert is changed among 30 minutes row (No.2) and the fresh 425 μ L TB (pH 6.5), comprises test substances is joined in the hole, top (giving body).Under 37 ℃,, in the polymix vibrator, plate is cultivated with the low jolting speed of about 150～300rpm.

Be expert at cultivated 30 minutes in 2 after, insert moved on in new pre-(receptor) hole, the outside, the end of heating up in per 30 minutes; Row 3 (60 minutes), 4 (90 minutes) and 5 (120 minutes).

After～2 minutes and when off-test, from the solution of summit, extract 25 μ L samples.These samples are represented on-test and are given the body sample when finishing.

At each preset time point, from (receptor) hole, the outside, the end, extract 300 μ L, and when off-test, measure the value afterwards of TEER.Adding acetonitrile in the sample of all collections, is 50% until its final concn in sample.The sample of all collections is stored under-20 ℃, till analyzing by HPLC or LC-MS.

The transhipment to the limit of the outside, the end

Usually, every kind of compound will be tested in 2～4 holes.The outside, the end and topped hole are 10 μ M with the normal concentration that contains 1.55mL and 0.4mL TB and test substances respectively.In addition, all test solns and damping fluid will contain 1%DMSO.Before test, make the transport plates precoating contain the developing medium 30 minutes of 10% serum, thereby avoid the non-specific combination with plastic material.

After 21～28 days, cell prepares for permeability test in the substratum on the screen support device.Developing medium is removed from topped hole and insert is changed in the washing row (No.1) of new plate (transport plates) of no insert.

This transport plates comprises 3 row, 4 holes of every row.Row 1 expression " washing " and row 3 are " sample is capable ".Transport plates is prepared by 1.5mL TB (pH 7.4) in washing row No.1 with by 1.55mL TB (pH 7.4) (comprising test substances) in test row No.3 (giving the side) in advance.

0.5mL transport buffer (pH 6.5 for HBSS, 25mM MES) is joined in the insert of capable No.1, and under 37 ℃, in the polymix vibrator, cell monolayer balance 30 minutes in the transhipment buffer system.After with the buffer system balance, the TEER value in each hole is measured by EVOM cutting adhesion instrument.

(TB, pH 6.5) removes from the top side with transport buffer, and changes insert over to row 3, and 400 fresh μ L TB (pH 6.5) are joined in the insert.After 30 minutes, from hole, top (receptor), take out 250 μ L and with fresh transport buffer replacement.After this, 250 μ L samples were taken out and replace in per 30 minutes with fresh transport buffer, until off-test in the 120th minute the time, the last value afterwards of when off-test, measuring TEER.After～2 minutes and when off-test, from the compartment of the outside, the end (giving body), take out 25 μ L samples.These samples are represented on-test and are given the body sample when finishing.

Adding acetonitrile in the sample of all collections, is 50% until its final concn in sample.The sample of all collections is stored under-20 ℃, till analyzing by HPLC or LC-MS.

Calculate

The running summary of the points scored FA that mensuration absorbed with respect to the time _CumFA _CumCalculate by following:

${\mathrm{FA}}_{\mathrm{cum}}=\mathrm{\&Sigma;}\frac{C_{\mathrm{RI}}}{C_{\mathrm{DI}}}$

C wherein _RiSusceptor concentration when finishing, and C for interval i _DiGive bulk concentration when beginning for interval i.Should obtain linear relationship.

Permeability coefficient (P _App, mensuration cm/s) is calculated by following:

$P_{\mathrm{app}}=\frac{(k\&CenterDot;V_R)}{(A\&CenterDot;60)}$

Wherein k is transport velocity (min ^-1), be defined as by being the running summary of the points scored (FA of the absorption of function with the time (min) _Cum) the slope that obtains of linear regression, V _RBe the area (cm that volume (mL) in the absorption chamber and A are strainer ²).

Reference compound

The reference that all are quoted in this application comprises patent and patent application, all is incorporated herein by reference on the maximum possible degree.

In whole specification sheets and claim subsequently, unless context has needs in addition, word " comprises " and variant, should be understood to comprise described integer, step, group of integers or step group such as " comprising " and " containing ", but do not repel any other integer, step, group of integers or step group.

Claims (30)

1. formula II compound

Wherein

R ¹And R ²In one of be that halogen and another are H or halogen;

R ³Be optional fluorizated-C ₁-C ₅Straight chain or branched-chain alkyl, perhaps-CH ₂CR ⁵C ₃-C ₄-cycloalkyl;

R ⁴Be H;

R ⁵Be H, C ₁-C ₂Alkyl, C ₁-C ₂Haloalkyl, hydroxyl, OC ₁-C ₂Alkyl, fluorine;

R ⁶Be monocycle stable, optional replacement or two ring carbocyclic ring or heterocycles, described or each ring has 4,5 or 6 annular atomses and 0～3 substituting group that is selected from the heteroatoms of S, O and N and wherein chooses wantonly comprises that 1～3 is selected from R ₇The member;

R ₇Be independently selected from halogen, oxo, nitrile, nitro, C ₁-C ₄Alkyl ,-XNRdRe ,-XNReR ⁸,-NReXR ⁸, NH ₂CO-, X-R ⁸, X-O-R ⁸, O-X-R ⁸, X-C (=O) R ⁸, X-(C=O) NRdR ⁸, X-NReC (=O) R ⁸, X-NH SO _mR ⁸, X-S (=O) _mR ⁸, X-C (=O) OR ⁸, X-NReC (=O) OR ⁸

R ⁸Be H, C independently ₁-C ₄Alkyl, C ₃-C ₆Cycloalkyl, pyrrolidyl, piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, indolinyl, pyranyl, sulfo-pyranyl, furyl, thienyl, pyrryl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, indyl, phenyl, they are chosen wantonly separately and are selected from R by maximum 3 ⁹The member replace;

R ⁹Be independently selected from hydroxyl, XR ¹⁰,-XNRdRe ,-XNReR ¹⁰,-NReC ₁-C ₄Alkyl R ¹⁰,-S (=O) _mRe, cyano group, carboxyl, oxo, C ₁-C ₄Alkyl, C ₁-C ₄Haloalkyl, C ₁-C ₄-alkoxyl group, C ₁-C ₄Alkyloyl, formamyl;

R ¹⁰Be C ₃-C ₆Cycloalkyl, pyrrolidyl, piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, indolinyl, pyranyl, sulfo-pyranyl, furyl, thienyl, pyrryl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, indyl, phenyl, they are optional separately by C ₁-C ₄Alkyl, halogen, hydroxyl, C ₁-C ₄Alkoxyl group, cyano group ,-S (=O) _mRe, C ₁-C ₄Haloalkyl replaces;

X is associative key or C independently ₁-C ₄Alkylidene group;

Ra is H, C independently ₁-C ₄Alkyl or CH ₃C (=O);

Rb is C ₁-C ₄Haloalkyl;

Rc is H, C ₁-C ₄Alkyl; Perhaps Rc and R ⁶And the carbon atom that they all connect forms altogether as R ⁶Defined carbocyclic ring or heterocycle;

Rd is H, C independently ₁-C ₄Alkyl or CH ₃C (=O);

Re is H, C independently ₁-C ₄Alkyl; Perhaps

Rd forms optional by R with the N atom that Re is connected with them altogether ⁹The morpholine, piperidines, piperazine or the pyrrolidine ring that replace;

M is 0,1 or 2 independently;

Perhaps its pharmacy acceptable salt, hydrate or N-oxide compound.

2. according to the compound of claim 1, wherein this stereochemistry is as shown in following part-structure:

3. according to the compound of claim 1, wherein this stereochemistry is as shown in following part-structure:

4. according to the compound of claim 1, wherein Rb be trifluoromethyl and this stereochemistry as shown in following part-structure:

5. according to the compound of claim 1, R wherein ²Be fluorine and R ¹Be H.

6. according to the compound of claim 1, R wherein ³Be C ₁-C ₄Branched-chain alkyl.

7. according to the compound of claim 6, R wherein ³Be isobutyl-.

8. according to the compound of claim 1, be H wherein at the Ra shown in the formula II.

9. according to the compound of claim 1, R wherein ⁶Be substituted-phenyl.

10. according to the compound of claim 9, wherein substituting group comprise-NRdRe ,-CH ₂NRdRe ,-NReR ⁹,-NReXR ⁹, C ₁-C ₄Straight chain or branched-chain alkyl or-O-R ⁹

11. according to the compound of claim 10, wherein substituting group comprises:

-NH-CH ₂Phenyl ,-NHCH ₂Pyridyl or-the NH-phenyl, wherein each phenyl or pyridyl ring are by C ₁-C ₄-alkyl ,-NRaRb ,-NRbR ⁸Perhaps-NRbC ₁-C ₄Alkyl R ⁸Replace.

12. according to the compound of claim 9, wherein substituting group comprises C ₃-C ₆Cycloalkyl, pyrrolidyl, piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, indolinyl, pyranyl, sulfo-pyranyl, furyl, thienyl, pyrryl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, indyl, phenyl, they are optional separately by R ⁹Replace.

13. compound according to claim 12, wherein substituting group is selected from indolinyl, pyranyl, sulfo-pyranyl, pyrryl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, indyl, and they are optional separately by R ⁹Replace.

14. according to the compound of claim 13, wherein substituting group is thiazolyl, 5-methyl-thiazolyl or thienyl, they are optional separately by R ⁹Replace.

15. according to the compound of claim 14, wherein substituting group is thiazole-4-base, 5-methylthiazol-4-base or thiophene-2-base, they are optional separately by R ⁹Replace.

16. compound according to claim 15, wherein thiazolyl, 5-methylthiazol base or thienyl by morpholinyl, morpholinyl methyl-, piperidyl, piperidino methyl-, piperazinyl, piperazinyl methyl substituted, described morpholinyl, morpholinyl methyl-, piperidyl, piperidino methyl-, arbitrary by C in the piperazinyl, piperazinyl methyl ₁-C ₃Alkyl, fluorine, difluoro or C ₁-C ₃Alkyl-O-C ₁-C ₃Alkyl-replacement.

17. according to the compound of claim 16, wherein the substituting group of thiazolyl, 5-methylthiazol base or thienyl is by methyl substituted piperidin-4-yl, by C ₁-C ₃The piperazinyl that alkyl or methyl oxygen base ethyl-carry out N-replaces, the piperidines-1-ylmethyl that perhaps is not substituted or is replaced by fluorine or difluoro 4--.

18. according to the compound of claim 10, wherein substituting group comprises optional by R ⁹The morpholine, piperidines or the piperazine ring that replace.

19., comprise piperidin-4-yl or N-piperazinyl, replaced by Ra N-, perhaps piperidines-1-the base that is replaced by-NRdRe 4-according to the compound of claim 18.

20. according to the compound of claim 1, wherein R ⁶Be benzothiazolyl, benzofuryl, 3-methyl benzofuryl or benzoxazolyl, they are optional separately by one or two R ⁷Replace.

21. according to the compound of claim 20, one of them described substituting group is-OR ⁸,-OXR ⁸,-NReR ⁸Perhaps-NReXR ⁸

22. according to the compound of claim 21, wherein R ⁸Be piperidin-4-yl, piperazine-1-base or piperidines-1-base or morpholino, they are arbitrary by C ₁-C ₃Alkyl replaces.

23. according to the compound of claim 22, wherein R ⁶Optional substituting group be N-morpholinyl oxyethyl group, N-morpholinyl methoxy base, N-methyl piperidine-4-base oxygen base or N-methylmorpholine-3-ylmethoxy.

24. according to the compound of claim 1, Ren Xuan substituent R wherein ⁹Be selected from hydroxyl, XR ¹⁰,-XNRdRe ,-XNReR ¹⁰,-NReC ₁-C ₄Alkyl R ¹⁰,-cyano group, carboxyl, oxo, C ₁-C ₄Alkyl, C ₁-C ₄-alkoxyl group, C ₁-C ₄Alkyloyl or formamyl.

25. a pharmaceutical composition comprises as each defined compound of claim 1～24 and pharmaceutically acceptable carrier or thinner.

26. as claim 1-24 in each defined compound be used for the treatment of purposes in the medicine of the inappropriate expression that is characterised in that cathepsin K or activatory disease in manufacturing.

27. according to the purposes of claim 26, wherein said disease is selected from:

Osteoporosis,

Gum disease, such as gingivitis and periodontitis,

Paget's disease,

The hypercalcemia of malignant tumour

The metabolism skeletal diseases

Be characterised in that the disease of excessive cartilage or matrix degraded, such as osteoarthritis and rheumatoid arthritis,

Osteocarcinoma comprises that tumour forms,

Pain.

28. as each defined compound of claim 1～24, as medicine.

29., be used for the treatment of or prevent to be characterised in that the inappropriate expression or the activatory disease of cathepsin K as each defined compound of claim 1-24.

30. treatment or prevention are characterised in that the inappropriate expression of cathepsin K or the method for activatory disease, comprise administration need its main body safety and significant quantity according to each compound of claim 1～24.